WO2015184001A1 - Glycoanticorps anti-thf-alpha et leurs utilisations - Google Patents

Glycoanticorps anti-thf-alpha et leurs utilisations Download PDF

Info

Publication number
WO2015184001A1
WO2015184001A1 PCT/US2015/032737 US2015032737W WO2015184001A1 WO 2015184001 A1 WO2015184001 A1 WO 2015184001A1 US 2015032737 W US2015032737 W US 2015032737W WO 2015184001 A1 WO2015184001 A1 WO 2015184001A1
Authority
WO
WIPO (PCT)
Prior art keywords
glcnac
sia
man
gal
tnfa
Prior art date
Application number
PCT/US2015/032737
Other languages
English (en)
Other versions
WO2015184001A8 (fr
Inventor
Chi-Heuy Wong
Chung-Yi Wu
Original Assignee
Academia Sinica
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Academia Sinica filed Critical Academia Sinica
Priority to CN201580027936.6A priority Critical patent/CN106714829A/zh
Priority to JP2016569732A priority patent/JP7063538B2/ja
Priority to CA2950433A priority patent/CA2950433A1/fr
Priority to EP15800059.6A priority patent/EP3154582A4/fr
Priority to KR1020167036499A priority patent/KR102494193B1/ko
Priority to AU2015267044A priority patent/AU2015267044A1/en
Publication of WO2015184001A1 publication Critical patent/WO2015184001A1/fr
Priority to IL249184A priority patent/IL249184B/en
Publication of WO2015184001A8 publication Critical patent/WO2015184001A8/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • C07K16/241—Tumor Necrosis Factors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/005—Glycopeptides, glycoproteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01051—Alpha-L-fucosidase (3.2.1.51)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A61K2039/507—Comprising a combination of two or more separate antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/72—Increased effector function due to an Fc-modification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • Fc glycosylation has been an important subject in the field of therapeutic monoclonal antibodies. Fc glycosylation can significantly modify Fc effector functions such as Fc receptor binding and complement activation, and thus affect the in vivo safety and efficacy profiles of therapeutic antibodies.
  • Non-human mammalian expression systems such as CHO or NS0 cells have the machinery required to add complex, human-type glycans.
  • glycans produced in these systems can differ from glycans produced in humans.
  • Their glycosylation machinery often adds undesired carbohydrate determinants which may alter protein folding, induce immunogenicity, and reduce circulatory life span of the drug.
  • sialic acid as N-acetylneuraminic acid is not efficiently added in most mammalian cells and the 6-linkage is missing in these cells.
  • TNFa Necrosis Factor Alpha
  • rheumatoid arthritis psoriatic arthritis
  • Ankylosing spondylitis Crohn's disease
  • psoriasis a major contributor to inflammation in autoimmune diseases, such as rheumatoid arthritis, psoriatic arthritis, Ankylosing spondylitis, Crohn's disease and psoriasis.
  • Adalimumab (Humira®) is a human-derived recombinant IgGl monoclonal antibody engineered by gene technology. Adalimumab binds to TNF-a but not TNF- ⁇ and has a half-life of approximately 2 weeks. It was approved for use in patients with RA December 31 , 2002.
  • Adalimumab is produced in Chinese hamster ovary (CHO) cells and are highly heterogeneous in glycosylation patterns in the Fc domains. Each of IgG molecules in the heterogeneous mixture may not all have the same property, and certain N-linked
  • oligosaccharides bound to therapeutic proteins may trigger undesired effects in patients thus deeming them a safety concern.
  • one aspect of the present disclosure relates to a composition of anti- TNFa glycoantibodies comprising a homogeneous population of anti-TNFa IgG molecules having the same N-glycan on each of Fc.
  • the anti-TNFa glycoantibodies of the invention can be produced from anti-TNFa monoclonal antibodies by Fc glycoengineering.
  • anti- TNFa glycoantibodies described herein have improved therapeutic values with increased TNFa binding affinity compared to the corresponding monoclonal antibodies that have not been glycoengineered.
  • the N-glycan is attached to the Asn-297 of the Fc region.
  • the anti-TNFa glycoantibody described herein comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 1, and a light chain having the amino acid sequence set forth in SEQ ID NO: 2.
  • the anti-TNFa glycoantibody described herein comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 1, and a light chain having the amino acid sequence set forth in SEQ ID NO: 2.
  • glycoantibody comprises a light chain sequence and a heavy chain sequence of Adalimumab (Humira®).
  • Humira® Adalimumab
  • the anti-TNFa glycoantibodies exhibit similar or improved TNFa binding affinity as compared to Adalimumab.
  • the N-glycan in the glycoantibody of the invention has a biantennary structure.
  • the N-glycan comprises a bisecting GlcNAc.
  • the N-glycan described herein comprises at least one a2-6 terminal sialic acid. In certain embodiments, the N-glycan comprises one a2-6 terminal sialic acid. In a preferred embodiment, the N-glycan comprises two a2-6 terminal sialic acids.
  • the N-glycan described herein comprises at least one a2-3 terminal sialic acid. In certain embodiments, the N-glycan comprises one a2-3 terminal sialic acid. In a preferred embodiment, the N-glycan comprises two a2-3 terminal sialic acids.
  • the N-glycan described herein comprises at least one galactose. In certain embodiments, the N-glycan comprises one galactose. In a preferred embodiment, the N-glycan comprises two galactoses.
  • the N-glycan is fucosylated. In some embodiments, the N- glycan is defucosylated.
  • the N-glycan has the glycan sequence selected from the group consisting of Sia2(a2-6)Gal 2 GlcNAc2Man 3 GlcNAc2, Sia(a2-6)Gal 2 GlcNAc2Man 3 GlcNAc2, Sia(a2-6)GalGlcNAc 2 Man 3 GlcNAc 2 , Sia 2 (a2-6)Gal 2 GlcNAc3Man3GlcNAc2, Sia(a2- 6)Gal 2 GlcNAc3Man 3 GlcNAc2, Sia(a2-6)GalGlcNAc 3 Man 3 GlcNAc 2 , Sia 2 (a2- 3)Gal 2 GlcNAc 2 Man 3 GlcNAc2, Sia (a2-3)GalGlcNAc 3 Man 3 GlcNAc 2 , Sia 2 (a2- 3)Gal 2 GlcNAc 3 Man 3 GlcNAc 2 , Sia 2 (a2- 3)Gal 2 GlcNA
  • the N-glycan has the glycan sequence selected from the group consisting of Sia 2 (a2-6)Gal 2 GlcNAc2Man 3 GlcNAc2, Sia(a2- 6)GalGlcNAc 2 Man 3 GlcNAc 2 , Sia 2 (a2-3)Gal 2 GlcNAc2Man 3 GlcNAc2, Sia2(a2- 3)Gal 2 GlcNAc 3 Man 3 GlcNAc2, Sia 2 (a2-3/a2-6)Gal 2 GlcNAc 3 Man 3 GlcNAc 2 , Sia 2 (a2-6/a2- 3)Gal 2 GlcNAc 3 Man 3 GlcNAc 2 , Sia 2 (a2-6)Gal 2 GlcNAc 2 Man 3 GlcNAc 2 (F), Sia(a2- 6)GalGlcNAc 2 Man 3 GlcNAc 2 (F), Sia 2 (a2-3)Gal 2 GlcNAc 2 (F), Sia(a
  • compositions comprising a composition of anti-TNFa glycoantibodies described herein and a
  • the pharmaceutical composition according to the disclosure may be used in therapeutics.
  • Disclosed herein include methods for treating a TNF-mediated inflammatory disease in a human patient, the method comprising administering a human in need thereof an effective amount of a pharmaceutical composition described herein.
  • the administering comprises the orally administering said pharmaceutical composition to said human.
  • TNF-mediated inflammatory diseases include, but not limited to, inflammatory bowel disease (including ulcerative colitis and Crohn's disease), rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, and juvenile idiopathic arthritis.
  • the treatment method described herein further comprises administering to the patient an anti-TNFa therapeutic agent.
  • the anti- TNFa therapeutic agent is Adalimumab, Infliximab or Etanercept.
  • the anti-TNFa glycoantibodies of the invention may be generated from anti-TNFa monoclonal antibodies produced in cells.
  • the anti-TNFa monoclonal antibodies are for therapeutic use.
  • the therapeutic monoclonal antibodies are commercially available or in development.
  • the anti-TNFa monoclonal antibodies can be humanized, human or chimeric.
  • the anti-TNFa glycoantibodies described herein can be produced in vitro.
  • the anti- TNFa glycoantibodies can be generated by Fc glycoengineering.
  • the anti-TNFa glycoantibodies are enzymatically or chemoenzymatically engineered from the anti- TNFa monoclonal antibodies produced in cells such as mammalian cells.
  • Described herein is a method for making a fucosylated glycoantibody, the method comprising the steps of (a) contacting a monoclonal antibody with at least one endoglycosidase, thereby yielding an antibody having a disaccharide (GlcNAc -Fuc) on the Fc, and (b) adding a carbohydrate moiety to GlcNAc of the disaccharide under suitable conditions.
  • Described herein also includes a method for making a defucosylated glycoantibody, the method comprising the steps of (a) contacting a monoclonal antibody with an a-fucosidase and at least one endoglycosidase, thereby yielding an antibody having a monosaccharide (GlcNAc) on the Fc, and (b) adding a carbohydrate moiety to GlcNAc under suitable conditions.
  • the monoclonal antibody in the methods of the invention may be an anti- TNFa monoclonal antibody.
  • the anti-TNFa monoclonal antibody in the methods of the invention is Adalimumab, Infliximab or Etanercept.
  • the carbohydrate moiety is selected from the group consisting of Sia 2 (a2-6)Gal 2 GlcNAc 2 Man 3 GlcNAc, Sia(a2-6)GalGlcNAc 2 Man 3 GlcNAc, Sia 2 (a2- 3)Gal 2 GlcNAc 2 Man 3 GlcNAc, Sia2(a2-3)Gal 2 GlcNAc 3 Man 3 GlcNAc, Sia 2 (a2-3/a2- 6)Gal 2 GlcNAc 3 Man 3 GlcNAc, Sia 2 (a2-6/a2-3)Gal 2 GlcNAc 3 Man 3 GlcNAc, Sia 2 (a2- 6)Gal 2 GlcNAc 2 Man 3 GlcNAc(F), Sia(a2-6)GalGlcNAc 2 Man 3 GlcNAc(F), Sia 2 (a2- 3)Gal 2 GlcNAc 2 Man 3 GlcNAc(F), Sia 2 (a2- 3)Gal 2
  • the adding in step (b) can be performed by a transglycosylase.
  • Transglycosylase includes, but are not limited to, EndoS, EndoH, EndoA, EndoM, EndoF l, EndoF2 and EndoF3.
  • the endoglycosidases in the method of the invention is EndoA, EndoF, EndoFl, EndoF2, EndoF3, EndoH, EndoM, EndoS or a variant thereof.
  • the alpha-fucosidase comprises a polypeptide having an amino acid sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical to SEQ ID NO: 5.
  • the alpha-fucosidase is a recombinant Bacteroides alpha-L- fucosidase.
  • the carbohydrate moiety is a sugar oxazoline.
  • Figure 1 SDS-PAGE analysis for anti-TNFa GAb 201 and lOl(upper), and GAb 200 and 401 (bottom). N-glycan profiling for anti-TNFa GAb 101 (upper), and GAb 401 (bottom).
  • Figure 2 shows binding affinity of TNFa to anti-TNFa GAb 200, 401 and
  • glycoantibodies The present disclosure relates to the development of a novel class of monoclonal antibodies, named "glycoantibodies".
  • the term "glycoantibodies” was coined by the inventor, Dr. Chi-Huey Wong, to refer to a homogeneous population of monoclonal antibodies (preferably, therapeutic monoclonal antibodies) having a single, uniform glycoform on Fc.
  • the individual glycoantibodies comprising the homogeneous population are identical, bind to the same epitope, and contain the same Fc glycan with a well-defined glycan structure and sequence.
  • anti-TNFa glycoantibodies refers to a homogeneous population of anti-TNFa IgG molecules having the same glycoform on Fc.
  • anti-TNFa glycoantibody refers to an individual IgG molecule in anti-TNFa glycoantibodies.
  • molecule can also refer to antigen binding fragments.
  • glycocan refers to a polysaccharide, oligosaccharide or monosaccharide. Glycans can be monomers or polymers of sugar residues and can be linear or branched.
  • a glycan may include natural sugar residues (e.g., glucose, N-acetylglucosamine, N- acetyl neuraminic acid, galactose, mannose, fucose, hexose, arabinose, ribose, xylose, etc.) and/or modified sugars (e.g., 2'-fluororibose, 2'-deoxyribose, phosphomannose, 6' sulfo N- acetylglucosamine, etc).
  • natural sugar residues e.g., glucose, N-acetylglucosamine, N- acetyl neuraminic acid, galactose, mannose, fucose, hexose, arabinose, ribose, xylose, etc.
  • modified sugars e.g., 2'-fluororibose, 2'-deoxyribose, phosphomannose,
  • Glycans can be monomers or polymers of sugar residues and can be linear or branched.
  • a glycan may include natural sugar residues (e.g., glucose, N- acetylglucosamine, N-acetyl neuraminic acid, galactose, mannose, fucose, hexose, arabinose, ribose, xylose, etc.) and/or modified sugars (e.g., 2'-fluororibose, 2'-deoxyribose,
  • natural sugar residues e.g., glucose, N- acetylglucosamine, N-acetyl neuraminic acid, galactose, mannose, fucose, hexose, arabinose, ribose, xylose, etc.
  • modified sugars e.g., 2'-fluororibose, 2'-deoxyribose
  • Glycan is also used herein to refer to the carbohydrate portion of a glycoconjugate, such as a glycoprotein, glycolipid, glycopeptide, glycoproteome, peptidoglycan, lipopolysaccharide or a proteoglycan.
  • Glycans usually consist solely of O-glycosidic linkages between monosaccharides.
  • cellulose is a glycan (or more specifically a glucan) composed of B-l,4-linked D-glucose
  • chitin is a glycan composed of B-l,4-linked N-acetyl-D-glucosamine.
  • Glycans can be homo or heteropolymers of monosaccharide residues, and can be linear or branched. Glycans can be found attached to proteins as in glycoproteins and proteoglycans. They are generally found on the exterior surface of cells. O- and N-linked glycans are very common in eukaryotes but may also be found, although less commonly, in prokaryotes. N-Linked glycans are found attached to the R-group nitrogen (N) of asparagine in the sequon. The sequon is a Asn-X-Ser or Asn-X-Thr sequence, where X is any amino acid except praline.
  • fucose As used herein, the terms “fucose”, “core fucose” and “core fucose residue” are used interchangeably and refer to a fucose in al,6-position linked to the N-acetylglucosamine .
  • fucose refers to the presence of a core fucose in the N-glycan of Fc
  • defucosylated refers to the absence of a core fucose in the N-glycan of Fc.
  • N-glycan As used herein, the terms "N-glycan”, “N-linked glycan”, “N-linked glycosylation”, “Fc glycan” and “Fc glycosylation” are used interchangeably and refer to an N-linked oligosaccharide attached by an N-acetylglucosamine (GlcNAc) linked to the amide nitrogen of an asparagine residue in a Fc-containing polypeptide.
  • GlcNAc N-acetylglucosamine
  • Fc-containing polypeptide refers to a polypeptide, such as an antibody, which comprises an Fc region.
  • glycosylation pattern and “glycosylation profile” are used interchangeably and refer to the characteristic "fingerprint” of the N-glycan species that have been released from a glycoprotein or antibody, either enzymatically or chemically, and then analyzed for their carbohydrate structure, for example, using LC-HPLC, or MALDI-TOF MS, and the like. See, for example, the review in Current Analytical Chemistry, Vol. 1, No. 1 (2005), pp. 28-57; herein incorporated by reference in its entirety.
  • glycoengineered Fc when used herein refers to N-glycan on the Fc region has been altered or engineered either enzymatically or chemically.
  • Fc glycoengineering refers to the enzymatic or chemical process used to make the glycoengineered Fc. Exemplary methods of engineering are described in, for example, Wong et al USSN12/959,351, the contents of which is hereby incorporated by reference.
  • glycan can be prepared by the use of endo-GlcNACase and fucosidase, then by the use of endo-S mutant and a glycan oxazoline.
  • effector function refers to a biochemical event that results from the interaction of an antibody Fc region with an Fc receptor or ligand.
  • exemplary "effector functions” include Clq binding; complement dependent cytotoxicity; Fc receptor binding;
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • phagocytosis phagocytosis
  • down regulation of cell surface receptors e.g. B cell receptor; BCR
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • FcRs Fc receptors
  • cytotoxic cells e.g. Natural Killer (NK) cells, neutrophils, and macrophages
  • NK cells Natural Killer cells
  • neutrophils neutrophils
  • macrophages cytotoxic cells
  • the antibodies “arm” the cytotoxic cells and are absolutely required for such killing.
  • the primary cells for mediating ADCC, NK cells express FcyRIII only, whereas monocytes express FcyRI, FcyRII and FcyRIII.
  • ADCC activity of a molecule of interest is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991).
  • an in vitro ADCC assay such as that described in U.S. Pat. No. 5,500,362 or U.S. Pat. No. 5,821,337 may be performed.
  • Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.
  • PBMC peripheral blood mononuclear cells
  • NK Natural Killer
  • ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).
  • CDC complement dependent cytotoxicity
  • Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (Clq) to antibodies (of the appropriate subclass) which are bound to their cognate antigen.
  • Clq first component of the complement system
  • a CDC assay e.g. as described in Gazzano-Santoro et al, J. Immunol. Methods 202: 163 (1996), may be performed.
  • An antibody that "induces apoptosis" is one which induces programmed cell death as determined by binding of annexin V, fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies).
  • the cell is an infected cell.
  • phosphatidyl serine (PS) translocation can be measured by annexin binding; DNA fragmentation can be evaluated through DNA laddering; and nuclear/chromatin condensation along with DNA fragmentation can be evaluated by any increase in hypodiploid cells.
  • PS phosphatidyl serine
  • the antibody that induces apoptosis is one that results in about 2 to 50 fold, preferably about 5 to 50 fold, and most preferably about 10 to 50 fold, induction of annexin binding relative to untreated cell in an annexin binding assay.
  • “Chimeric" antibodies have a portion of the heavy and/or light chain identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81 :6851-6855 (1984)).
  • Humanized antibody as used herein is a subset of chimeric antibodies.
  • Humanized forms of non-human (e.g., murine) antibodies are chimeric antibodies which contain minimal sequence derived from non-human immunoglobulin.
  • humanized antibodies are human immunoglobulins (recipient or acceptor antibody) in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • donor antibody such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • humanized antibodies may comprise residues which are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance such as binding affinity.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence although the FR regions may include one or more amino acid substitutions that improve binding affinity.
  • the number of these amino acid substitutions in the FR is typically no more than 6 in the H chain, and in the L chain, no more than 3.
  • the humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human
  • Treating” or “treatment” or “alleviation” refers to both therapeutic treatment and prophylactic or preventative measures; wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder.
  • Those in need of treatment include those already with the disorder as well as those prone to have the disorder or those in whom the disorder is to be prevented.
  • a subject or mammal is successfully "treated" for an infection if, after receiving a therapeutic amount of an antibody according to the methods of the present invention, the patient shows observable and/or measurable reduction in or absence of one or more of the following: reduction in the number of infected cells or absence of the infected cells; reduction in the percent of total cells that are infected; and/or relief to some extent, one or more of the symptoms associated with the specific infection; reduced morbidity and mortality, and improvement in quality of life issues.
  • the above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician.
  • Administration "in combination with” one or more further therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order.
  • Carriers as used herein include pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution.
  • physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or nonionic surfactants such as TWEENTM polyethylene glycol (PEG), and PLURONICSTM
  • glycoantibodies was coined by the inventor, Dr. Chi-Huey Wong, to refer to a homogeneous population of monoclonal antibodies (preferably, therapeutic monoclonal antibodies) having a single, uniformed glycoform bound to the Fc region.
  • the individual glycoantibodies comprising the essentially homogeneous population are identical, bind to the same epitope, and contain the same Fc glycan with a well-defined glycan structure and sequence.
  • homogeneous in the context of a glycosylation profile of Fc region are used interchangeably and are intended to mean a single glycosylation pattern represented by one desired N-glycan species, with little or no trace amount of precursor N-glycan.
  • the trace amount of the precursor N- glycan is less than about 2%.
  • Essentially pure protein means a composition comprising at least about 90% by weight of the protein, based on total weight of the composition, including, for example, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% by weight.
  • Essentially homogeneous protein means a composition comprising at least about 98% by weight of protein, including for example, at least about 98.5 %, at least about 99% based on total weight of the composition.
  • the protein is an antibody, structural variants, and/or antigen binding fragment thereof.
  • immunoglobulin and “immunoglobulin molecule” are used interchangeably.
  • molecule can also refer to antigen binding fragments.
  • Fc receptor or "FcR” describes a receptor that binds to the Fc region of an antibody.
  • the preferred FcR is a native sequence human FcR.
  • a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcyRI (CD64), FcyRII (CD32), and FcyRIII (CD 16) subclasses, including allelic variants and alternatively spliced forms of these receptors.
  • FcyRII receptors include FcyRIIA (an “activating receptor”) and FcyRIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof.
  • Activating receptor FcyRIIA contains an immunoreceptor tyrosine-based activation motif (IT AM) in its cytoplasmic domain.
  • Inhibiting receptor FcyRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain, (see review M. in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)).
  • FcRs are reviewed in Ravetch and Kinet, Annu. Rev.
  • FcR neonatal receptor
  • the term "antigen” is defined as any substance capable of eliciting an immune response.
  • the term “antigen specific” refers to a property of a cell population such that supply of a particular antigen, or a fragment of the antigen, results in specific cell proliferation.
  • immunoogenicity refers to the ability of an
  • immunogen antigen, or vaccine to stimulate an immune response.
  • epitope is defined as the parts of an antigen molecule which contact the antigen binding site of an antibody or a T cell receptor.
  • specifically binding refers to the interaction between binding pairs (e.g., an antibody and an antigen). In various instances, specifically binding can be embodied by an affinity constant of about 10-6 moles/liter, about 10-7 moles/liter, or about 10-8 moles/liter, or less.
  • An "isolated" antibody is one which has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials which would interfere with research, diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or
  • substantially equivalent denotes a sufficiently high degree of similarity between two numeric values (for example, one associated with a molecule and the other associated with a reference/comparator molecule) such that one of skill in the art would consider the difference between the two values to be of little or no biological and/or statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values, anti-viral effects, etc.).
  • the difference between said two values is, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and/or less than about 10% as a function of the value for the reference/comparator molecule.
  • the phrase "substantially reduced,” or “substantially different”, as used herein, denotes a sufficiently high degree of difference between two numeric values (generally one associated with a molecule and the other associated with a reference/comparator molecule) such that one of skill in the art would consider the difference between the two values to be of statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values).
  • the difference between said two values is, for example, greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, and/or greater than about 50% as a function of the value for the reference/comparator molecule.
  • Binding affinity generally refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, as used herein, "binding affinity” refers to intrinsic binding affinity which reflects a 1 : 1 interaction between members of a binding pair (e.g., antibody and antigen).
  • the affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein.
  • Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigen faster and tend to remain bound longer.
  • a variety of methods of measuring binding affinity are known in the art, any of which can be used for purposes of the present invention. Specific illustrative embodiments are described in the following.
  • variable region or “variable domain” of an antibody refers to the amino- terminal domains of heavy or light chain of the antibody. These domains are generally the most variable parts of an antibody and contain the antigen-binding sites.
  • variable refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called complementarity-determining regions (CDRs) or hypervariable regions both in the light- chain and the heavy-chain variable domains. The more highly conserved portions of variable domains are called the framework (FR).
  • CDRs complementarity-determining regions
  • FR framework
  • the variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure.
  • the CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al, Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)).
  • the constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.
  • Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen-binding site, and a residual "Fc” fragment, whose name reflects its ability to crystallize readily.
  • Pepsin treatment yields an F(ab')2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen.
  • Fv is the minimum antibody fragment which contains a complete antigen- recognition and -binding site.
  • this region consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association.
  • one heavy- and one light-chain variable domain can be covalently linked by a flexible peptide linker such that the light and heavy chains can associate in a "dimeric" structure analogous to that in a two-chain Fv species. It is in this configuration that the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer.
  • the six CDRs confer antigen-binding specificity to the antibody.
  • a single variable domain or half of an Fv comprising only three CDRs specific for an antigen
  • the Fab fragment also contains the constant domain of the light chain and the first constant domain (CHI) of the heavy chain.
  • Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region.
  • Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group.
  • F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.
  • the "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa ( ⁇ ) and lambda ( ⁇ ), based on the amino acid sequences of their constant domains.
  • antibodies (immunoglobulins) can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.
  • the heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, ⁇ , ⁇ , ⁇ , and ⁇ , respectively.
  • the subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and described generally in, for example, Abbas et al. Cellular and Mol. Immunology, 4th ed. (2000).
  • An antibody may be part of a larger fusion molecule, formed by covalent or non-covalent association of the antibody with one or more other proteins or peptides.
  • full length antibody intact antibody
  • whole antibody are used herein interchangeably, to refer to an antibody in its substantially intact form, not antibody fragments as defined below.
  • Antibody fragments comprise only a portion of an intact antibody, wherein the portion retains at least one, and as many as most or all, of the functions normally associated with that portion when present in an intact antibody.
  • an antibody fragment comprises an antigen binding site of the intact antibody and thus retains the ability to bind antigen.
  • an antibody fragment for example one that comprises the Fc region, retains at least one of the biological functions normally associated with the Fc region when present in an intact antibody, such as FcRn binding, antibody half life modulation, ADCC function and complement binding.
  • an antibody fragment is a monovalent antibody that has an in vivo half life substantially similar to an intact antibody.
  • such an antibody fragment may comprise an antigen binding arm linked to an Fc sequence capable of conferring in vivo stability to the fragment.
  • the term "monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Thus, the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies.
  • Such monoclonal antibody typically includes an antibody comprising a polypeptide sequence that binds a target, wherein the target- binding polypeptide sequence was obtained by a process that includes the selection of a single target binding polypeptide sequence from a plurality of polypeptide sequences.
  • the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones or recombinant DNA clones.
  • the selected target binding sequence can be further altered, for example, to improve affinity for the target, to humanize the target binding sequence, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc., and that an antibody comprising the altered target binding sequence is also a monoclonal antibody of this invention.
  • each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen.
  • the monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.
  • the modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies to be used in accordance with the present invention may be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler et al, Nature, 256: 495 (1975); Harlow et al, Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al, in: Monoclonal Antibodies and T- Cell hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No.
  • phage display technologies See, e.g., Clackson et al, Nature, 352: 624-628 (1991); Marks et al, J. Mol. Biol. 222: 581-597 (1992); Sidhu et al, J. Mol. Biol. 338(2): 299- 310 (2004); Lee et al, J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al, J. Immunol.
  • the monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al, Proc. Natl. Acad. Sci. USA 81 :6851-6855 (1984)).
  • hypervariable region when used herein refers to the regions of an antibody variable domain which are hypervariable in sequence and/or form structurally defined loops.
  • antibodies comprise six hypervariable regions; three in the VH (HI, H2, H3), and three in the VL (LI, L2, L3).
  • a number of hypervariable region delineations are in use and are encompassed herein.
  • the Kabat Complementarity Determining Regions are based on sequence variability and are the most commonly used (Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).
  • Chothia refers instead to the location of the structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)).
  • the AbM hypervariable regions represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular's AbM antibody modeling software.
  • the "contact" hypervariable regions are based on an analysis of the available complex crystal structures. The residues from each of these hypervariable regions are noted below.
  • Hypervariable regions may comprise "extended hypervariable regions” as follows: 24-36 or 24-34 (LI), 46-56 or 50-56 or 49-56 (L2) and 89-97 or 89-96 (L3) in the VL and 26-35 (HI), 50-65 or 49-65 (H2) and 93-102, 94-102, or 95-102 (H3) in the VH.
  • the variable domain residues are numbered according to Kabat et al, supra, for each of these definitions.
  • “Framework” or "FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
  • variable domain residue numbering as in Kabat or "amino acid position numbering as in Kabat,” and variations thereof, refers to the numbering system used for heavy chain variable domains or light chain variable domains of the compilation of antibodies in Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a FR or HVR of the variable domain.
  • a heavy chain variable domain may include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g. residues 82a, 82b, and 82c, etc. according to Kabat) after heavy chain FR residue 82.
  • the Kabat numbering of residues may be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a "standard" Kabat numbered sequence.
  • Single-chain Fv or “scFv” antibody fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain.
  • the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding.
  • a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding.
  • diabodies refers to small antibody fragments with two antigen- binding sites, which fragments comprise a heavy -chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL).
  • VH heavy -chain variable domain
  • VL light-chain variable domain
  • a "human antibody” is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and/or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues.
  • an "affinity matured" antibody is one with one or more alterations in one or more HVRs thereof which result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess those alteration(s).
  • an affinity matured antibody has nanomolar or even picomolar affinities for the target antigen.
  • Affinity matured antibodies are produced by procedures known in the art. Marks et al.
  • a “blocking” antibody or an “antagonist” antibody is one which inhibits or reduces biological activity of the antigen it binds. Certain blocking antibodies or antagonist antibodies substantially or completely inhibit the biological activity of the antigen.
  • An "agonist antibody”, as used herein, is an antibody which mimics at least one of the functional activities of a polypeptide of interest.
  • a “disorder” is any condition that would benefit from treatment with an antibody of the invention. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question.
  • disorders to be treated herein include cancer.
  • cell proliferative disorder and “proliferative disorder” refer to disorders that are associated with some degree of abnormal cell proliferation. In one
  • the cell proliferative disorder is cancer.
  • Tumor refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues.
  • cancer refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues.
  • cancer refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues.
  • cancer cancer
  • cancer cancer
  • cancer cancer
  • cancer cancer
  • cancer cancer
  • cancer cancer
  • cancer and “cancerous” generally refer to or describe the
  • cancer physiological condition in mammals that is typically characterized by unregulated cell growth/proliferation.
  • cancer include, but are not limited to, carcinoma, lymphoma (e.g., Hodgkin's and non-Hodgkin's lymphoma), blastoma, sarcoma, and leukemia.
  • cancers include squamous cell cancer, small-cell lung cancer, non- small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, leukemia and other lymphoproliferative disorders, and various types of head and neck cancer.
  • the term "antigen" is defined as any substance capable of eliciting an immune response.
  • the term "antigen specific” refers to a property of a cell population such that supply of a particular antigen, or a fragment of the antigen, results in specific cell proliferation.
  • treatment refers to clinical intervention in an attempt to alter the natural course of the individual or cell being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing or decreasing
  • antibodies of the invention are used to delay development of a disease or disorder.
  • An "individual” or a “subject” is a vertebrate.
  • the vertebrate is a mammal. Mammals include, but are not limited to, farm animals (such as cows), sport animals, pets (such as cats, dogs, and horses), primates, mice and rats.
  • the vertebrate is a human.
  • "Mammal" for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, cows, etc. In certain embodiments, the mammal is human.
  • an “effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.
  • a "therapeutically effective amount" of a substance/molecule of the invention may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance/molecule, to elicit a desired response in the individual.
  • a therapeutically effective amount is also one in which any toxic or detrimental effects of the substance/molecule are outweighed by the therapeutically beneficial effects.
  • a “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically but not necessarily, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount would be less than the therapeutically effective amount.
  • cytotoxic agent refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells.
  • the term is intended to include radioactive isotopes (e.g., At211, 1131, 1125, Y90, Rel86, Rel 88, Sml53, Bi212, P32, Pb212 and radioactive isotopes of Lu), chemotherapeutic agents (e.g., methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents, enzymes and fragments thereof such as nucleolyticenzymes, antibiotics, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof, and the various antitum
  • a "chemotherapeutic agent” is a chemical compound useful in the treatment of cancer.
  • chemotherapeutic agents include alkylating agents such as thiotepa and CYTOXAN® cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARTNOL®); beta-lapachone; lapachol; colchicines; betulinic acid; a camptothecin (including the synthetic analogue
  • dynemicin including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® doxorubicin (including morpholino-doxorubicin, cyanomorpholino- doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mito
  • spirogermanium spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C”); thiotepa; taxoids, e.g., TAXOL® paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANETM Cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, 111.), and
  • TAXOTERE® doxetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil; gemcitabine (GEMZAR®); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine (VELBAN®); platinum; etoposide (VP- 16); ifosfamide;
  • NAVELBINE® novantrone
  • edatrexate daunomycin
  • aminopterin ibandronate
  • topoisomerase inhibitor RFS 2000 difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine (XELODA®); pharmaceutically acceptable salts, acids or derivatives of any of the above; as well as combinations of two or more of the above such as CHOP, an
  • treatment refers to clinical intervention in an attempt to alter the natural course of the individual or cell being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing or decreasing
  • antibodies of the invention are used to delay development of a disease or disorder.
  • an "individual” or a “subject” is a vertebrate.
  • the vertebrate is a mammal. Mammals include, but are not limited to, farm animals (such as cows), sport animals, pets (such as cats, dogs, and horses), primates, mice and rats.
  • the vertebrate is a human.
  • mammal for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, cows, etc. In certain embodiments, the mammal is human.
  • an “effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.
  • a “therapeutically effective amount” of a substance/molecule of the invention may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance/molecule, to elicit a desired response in the individual.
  • a therapeutically effective amount is also one in which any toxic or detrimental effects of the substance/molecule are outweighed by the therapeutically beneficial effects.
  • a “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically but not necessarily, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount would be less than the therapeutically effective amount.
  • cytotoxic agent refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells.
  • the term is intended to include radioactive isotopes (e.g., At211, 1131, 1125, Y90, Rel86, Rel 88, Sml53, Bi212, P32, Pb212 and radioactive isotopes of Lu), chemotherapeutic agents (e.g., methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents, enzymes and fragments thereof such as nucleolyticenzymes, antibiotics, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof, and the various antitum
  • Administration "in combination with” one or more further therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order.
  • glycosylation of recombinant proteins produced from mammalian cells in culture is an important process in ensuring the effective use of therapeutic antibodies (Goochee et al, 1991 ; Jenkins and Curling, 1994).
  • Mammalian cell culture delivers a heterogeneous mixture of glycosylation patterns which do not all have the same properties. Properties like safety, efficacy and the serum half-life of therapeutic proteins can be affected by these glycosylation patterns.
  • glycoform heterogeneity problem by the development of a novel class of monoclonal antibodies, named "glycoantibodies”.
  • glycoantibodies was coined by the inventor, Dr. Chi-Huey Wong, to refer to a homogeneous population of monoclonal antibodies (preferably, therapeutic monoclonal antibodies) having a single, uniformed glycoform on Fc.
  • the individual glycoantibodies comprising the homogeneous population are identical, bind to the same epitope, and contain the same Fc glycan with a well-defined glycan structure and sequence.
  • Glycoantibodies may be generated from monoclonal antibodies, preferably therapeutic monoclonal antibodies, produced in cells. Glycoantibodies may be commercially available or in the clinical development. Monoclonal antibodies for therapeutic use can be humanized, human or chimeric.
  • Examples of approved monoclonal antibodies for therapeutic use include, but not limited to, Muromomab, Abciximab, Rituximab, Daclizumab, Basiliximab, Palivizumab, Infliximab,Trastuzumab, Etanercept, Gemtuzumab, Alemtuzumab, Ibritomomab, Adalimumab, Alefacept, Omalizumab, Efalizumab, Cetuximab, Bevacizumab, Natalizumab, Ranibizumab, Panitumumab, Eculizumab and Certolizumab.
  • glycoantibodies derived from therapeutic monoclonal antibodies by Fc glycoengineering exhibit similar or better activities as compared to the therapeutic monoclonal antibodies. It is contemplated that the glycoantibodies with optimized glycoforms may provide an alternative for therapeutic use.
  • TNFa tumor necrosis factor-a
  • TNFp tumor necrosis factor- ⁇
  • TNFa is a soluble homotrimer of 17 kD protein subunits (Smith, et al, J. Biol. Chem. 262:6951-6954 (1987)).
  • a membrane-bound 26 kD precursor form of TNF also exists (Kriegler, et al, Cell 53 :45-53 (1988)).
  • TNF-a is a potent inducer of the inflammatory response, a key regulator of innate immunity and plays an important role in the regulation of Thl immune responses against intracellular bacteria and certain viral infections.
  • dysregulated TNF can also contribute to numerous pathological situations. These include immune-mediated inflammatory diseases (IMIDs) including rheumatoid arthritis, Crohn's disease, psoriatic arthritis, ankylosing spondylitis, ulcerative colitis and severe chronic plaque psoriasis.
  • IMIDs immune-mediated inflammatory diseases
  • anti-TNFa glycoantibodies termed "anti-TNFa GAbs"
  • Anti-TNFa glycoantibodies can be generated from anti-TNFa monoclonal antibodies ("parental antibodies”) by Fc
  • parental antibodies refers to the anti-TNFa monoclonal antibodies used to produce anti-TNFa glycoantibodies.
  • the individual anti- TNFa glycoantibodies comprising the homogeneous population are identical and contain the same Fc glycan with a well-defined glycan structure and sequence.
  • Anti-TNFa glycoantibodies of the invention may bind to the same epitope of a human TNFa antigen as its patental antibodies do.
  • the parental antibodies may be produced in cells such as mammalian cells, Pichia pastoris or insect cells. Preferrably, the parental antibodies are produced in mammalian cells.
  • the parental antibodies may be FDA approved or in development.
  • Anti-TNFa monoclonal antibodies approved or in development include Infliximab, Adalimumab, Golimumab, CDP870 (certolizumab), TNF-TeAb and CDP571.
  • An anti-TNFa glycoantibody of the invention may comprise a heavy chain having the amino acid sequence set forth in SEQ ID NO: 1, and a light chain having the amino acid sequence set forth in SEQ ID NO: 2.
  • An anti-TNFa glycoantibody of the invention may comprise a light chain sequence and a heavy chain sequence of Adalimumab (Humira®).
  • Table 1 below shows the heavy chain and the light chain sequences of Adalimumab. Table 1.
  • the N-glycan may be attached to the Asn-297 of the Fc region.
  • the N-glycans according to the invention have a common pentasaccharide core of Man 3 GlcNAc 2 which is also referred to as "trimannose core” or "pentasaccharide core", wherein “Man” refers to mannose, “Glc” refers to glucose, “NAc” refers to N-acetyl, and GlcNAc refers to N-acetylglucosamine.
  • the N-glycan described herein may have a biantennary structure.
  • the N-glycan may have intrachain substitutions comprising "bisecting" GlcNAc.
  • a glycan comprises a bisecting GlcNAc on the trimannose core
  • the structure is represented as Man 3 GlcNAc 3
  • a glycan comprises a core fucose attached to the trimannose core
  • the structure is represented as Man 3 GlcNAc 2 (F).
  • the N-glycan may comprise one or more termial sialic acids (e.g. N-acetylneuraminic acid).
  • the structure represented as "Sia" refers to a termial sialic acid. Sialylation may occur on either the al-3 or al-6 arm of the biantennary structures.
  • the N-glycan described herein may comprise at least one a2-6 terminal sialic acid.
  • the N-glycan comprises one a2-6 terminal sialic acid.
  • the N-glycan comprises two a2-6 terminal sialic acids.
  • the N-glycan may comprise at least one a2-3 terminal sialic acid.
  • the N-glycan comprises one a2-3 terminal sialic acid.
  • the N-glycan comprises two a2-3 terminal sialic acids.
  • the N-glycan may comprise at least one galactose. In certain embodiments, the N- glycan comprises one galactose. In a preferred embodiment, the N-glycan comprises two galactoses.
  • the N-glycan may be fucosylated or defucosylated. Preferrably, the N-glycan is fucosylated.
  • Table 2 lists exemplary N-glycans in anti-TNFa glycoantibodies. Embodiments of the present disclosure may include or exclude any of the N-glycans listed herein.
  • An anti-TNFa glycoantibody of the invention can be produced by Fc
  • the parental antibody is Adalimumab (Humira®).
  • Described herein is a method for making a fucosylated glycoantibody, the method comprising the steps of (a) contacting a monoclonal antibody with at least one endoglycosidase, thereby yielding an antibody having a disaccharide (GlcNAc -Fuc) on the Fc, and (b) adding a carbohydrate moiety to GlcNAc of the disaccharide under suitable conditions.
  • the monoclonal antibody in the method of the invention may be an anti- TNFa monoclonal antibody.
  • the anti-TNFa monoclonal antibody is Adalimumab, Infliximab or Etanercept.
  • the endoglycosidase used in the method of the invention may be an EndoA, EndoF, EndoFl, EndoF2, EndoF3, EndoH, EndoM, EndoS or a variant thereof.
  • the subsequent enzyme-mediated glycosylation in step (b) using a transglycosylase is performed by adding a carbohydrate moiety to GlcNAc to extend the sugar chain.
  • transglycosylases include, but not limited to, EndoA, EndoF, EndoFl, EndoF2, EndoF3, EndoH, EndoM, EndoS, and variants thereof.
  • glycosylation using a sugar oxazoline as the sugar donor is useful for synthesizing oligosaccharides because the glycosylation reaction is an addition reaction and advances without any accompanying elimination of acid, water, or the like (Fujita, et al, Biochim. Biophys. Acta 2001, 1528, 9-14).
  • the carbohydrate moiety is a sugar oxazoline.
  • the carbohydrate moiety is selected from the group consisting of Sia 2 (a2-6)Gal 2 GlcNAc 2 Man 3 GlcNAc, Sia(a2-6)GalGlcNAc 2 Man 3 GlcNAc, Sia 2 (a2- 3)Gal 2 GlcNAc 2 Man 3 GlcNAc, Sia2(a2-3)Gal 2 GlcNAc 3 Man 3 GlcNAc, Sia 2 (a2-3/a2- 6)Gal 2 GlcNAc 3 Man 3 GlcNAc, Sia 2 (a2-6/a2-3)Gal 2 GlcNAc 3 Man 3 GlcNAc, Sia 2 (a2- 6)Gal 2 GlcNAc 2 Man 3 GlcNAc(F), Sia(a2-6)GalGlcNAc 2 Man 3 GlcNAc(F), Sia 2 (a2- 3)Gal 2 GlcNAc 2 Man 3 GlcNAc(F), Sia 2 (a2- 3)Gal 2
  • Described herein also includes an improved method for making a defucosylated glycoantibody, the method comprising the steps of (a) contacting a monoclonal antibody with an a-fucosidase and at least one endoglycosidase, thereby yielding an antibody having a monosaccharide (GlcNAc) on the Fc, and (b) adding a carbohydrate moiety to GlcNAc under suitable conditions.
  • a monoclonal antibody with an a-fucosidase and at least one endoglycosidase
  • the monoclonal antibody according to the improved method of the invention may be an anti-TNFa monoclonal antibody.
  • the anti-TNFa monoclonal antibody is Adalimumab.
  • the a-fucosidase in the improved method of the invention comprises a polypeptide containing an amino acid sequence having at least 85% identity to the sequences of SEQ ID NO: 5, a functional variant thereof.
  • the a-fucosidase may comprise a polypeptide comprising an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the sequences of SEQ ID NO: 5 (listed in Table 3), a variant or a fragment thereof.
  • the ⁇ -fucosidase is a recombinant Bacteroides a-fucosidase.
  • the polypeptide of the a-fucosidase of the invention may be derivatized or modified to assist with their isolation or purification.
  • the polypeptide for use in the invention is derivatized or modified by addition of a ligand which is capable of binding directly and specifically to a separation means.
  • the polypeptide is derivatized or modified by addition of one member of a binding pair and the separation means comprises a reagent that is derivatized or modified by addition of the other member of a binding pair. Any suitable binding pair can be used.
  • the polypeptide for use in the invention is derivatized or modified by addition of one member of a binding pair
  • the polypeptide is preferably histidine-tagged or biotin-tagged.
  • the amino acid coding sequence of the histidine or biotin tag is included at the gene level and the proteins are expressed recombinantly in E. coli.
  • the histidine or biotin tag is typically present at one end of the polypeptide, either at the N-terminus or at the C-terminus.
  • the histidine tag typically consists of six histidine residues, although it can be longer than this, typically up to 7, 8, 9, 10 or 20 amino acids or shorter, for example 5, 4, 3, 2 or 1 amino acids.
  • the histidine tag may contain one or more amino acid substitutions, preferably conservative substitutions as defined above.
  • Variant polypeptide as described herein are those for which the amino acid sequence varies from that in SEQ ID NO: 5, but exhibit the same or similar function of the enzyme comprising the polypeptide having an amino acid sequence of SEQ ID NO: 5.
  • percent (%) sequence identity with respect to a sequence is defined as the percentage of amino acid residues in a candidate polypeptide sequence that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.
  • a "pre-lyophilized formulation” can be produced.
  • the anti-TNFa GAb for preparing the formulation is preferably essentially pure and desirably essentially homogeneous (i.e. free from contaminating proteins etc).
  • "Essentially pure” protein means a composition comprising at least about 90% by weight of the protein, based on total weight of the composition, preferably at least about 95% by weight.
  • "Essentially homogeneous” protein means a composition comprising at least about 99% by weight of protein, based on total weight of the composition.
  • the protein is an antibody.
  • the amount of anti-TNFa GAb in the pre-lyophilized formulation is determined taking into account the desired dose volumes, mode(s) of administration etc.
  • the protein of choice is an intact antibody (a full-length antibody)
  • from about 2 mg/mL to about 50 mg/mL, preferably from about 5 mg/mL to about 40 mg/mL and most preferably from about 20-30 mg/mL is an exemplary starting protein concentration.
  • the protein is generally present in solution.
  • the protein may be present in a pH-buffered solution at a pH from about 4-8, and preferably from about 5-7.
  • Exemplary buffers include histidine, phosphate, Tris, citrate, succinate and other organic acids.
  • the buffer concentration can be from about 1 mM to about 20 niM, or from about 3 mM to about 15 mM, depending, for example, on the buffer and the desired isotonicity of the formulation (e.g. of the reconstituted formulation).
  • the preferred buffer is histidine in that, as demonstrated below, this can have lyoprotective properties.
  • Succinate was shown to be another useful buffer.
  • the lyoprotectant is added to the pre-lyophilized formulation.
  • the lyoprotectant is a non-reducing sugar such as sucrose or trehalose.
  • the amount of lyoprotectant in the pre-lyophilized formulation is generally such that, upon reconstitution, the resulting formulation will be isotonic. However, hypertonic reconstituted formulations may also be suitable. In addition, the amount of lyoprotectant must not be too low such that an unacceptable amount of degradation/aggregation of the protein occurs upon lyophilization.
  • lyoprotectant concentrations in the pre-lyophilized formulation are from about 10 mM to about 400 mM, and preferably from about 30 mM to about 300 mM, and most preferably from about 50 mM to about 100 mM.
  • the ratio of protein to lyoprotectant is selected for each protein and lyoprotectant combination.
  • the molar ratio of lyoprotectant to antibody may be from about 100 to about 1500 moles lyoprotectant to 1 mole antibody, and preferably from about 200 to about 1000 moles of lyoprotectant to 1 mole antibody, for example from about 200 to about 600 moles of lyoprotectant to 1 mole antibody.
  • a surfactant to the pre-lyophilized formulation.
  • the surfactant may be added to the lyophilized formulation and/or the reconstituted formulation.
  • exemplary surfactants include nonionic surfactants such as polysorbates (e.g. polysorbates 20 or 80); poloxamers (e.g.
  • poloxamer 188 Triton; sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl- or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palnidopropyl-, or isostearamidopropyl-betaine (e.g lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl- dimethylamine; sodium methyl cocoyl
  • MONAQUATTM series Mona Industries, Inc., Paterson, N.J.
  • polyethyl glycol polypropyl glycol
  • copolymers of ethylene and propylene glycol e.g. Pluronics, PF68 etc.
  • the amount of surfactant added is such that it reduces aggregation of the reconstituted protein and minimizes the formation of particulates after reconstitution.
  • the surfactant may be present in the pre-lyophilized formulation in an amount from about 0.001-0.5%, and preferably from about 0.005-0.05%.
  • a mixture of the lyoprotectant such as sucrose or trehalose
  • a bulking agent e.g. mannitol or glycine
  • the bulking agent may allow for the production of a uniform lyophilized cake without excessive pockets therein etc.
  • compositions such as those described in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980) may be included in the pre-lyophilized formulation (and/or the lyophilized formulation and/or the reconstituted formulation) provided that they do not adversely affect the desired characteristics of the formulation.
  • Acceptable carriers, excipients or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include; additional buffering agents; preservatives; co-solvents; antioxidants including ascorbic acid and methionine; chelating agents such as EDTA; metal complexes (e.g. Zn-protein complexes); biodegradable polymers such as polyesters; and/or salt-forming counterions such as sodium.
  • compositions and formulations described herein are preferably stable.
  • a “stable" formulation/composition is one in which the antibody therein essentially retains its physical and chemical stability and integrity upon storage.
  • Various analytical techniques for measuring protein stability are available in the art and are reviewed in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993). Stability can be measured at a selected temperature for a selected time period.
  • the formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes, prior to, or following, lyophilization and reconstitution. Alternatively, sterility of the entire mixture may be accomplished by autoclaving the ingredients, except for protein, at about 120° C. for about 30 minutes, for example.
  • the formulation is lyophilized.
  • freeze-dryers are available for this purpose such as Hull50® (Hull, USA) or GT20® (Leybold-Heraeus, Germany) freeze-dryers. Freeze-drying is accomplished by freezing the formulation and subsequently subliming ice from the frozen content at a temperature suitable for primary drying. Under this condition, the product temperature is below the eutectic point or the collapse temperature of the formulation.
  • the shelf temperature for the primary drying will range from about -30 to 25° C. (provided the product remains frozen during primary drying) at a suitable pressure, ranging typically from about 50 to 250 mTorr.
  • the formulation, size and type of the container holding the sample (e.g., glass vial) and the volume of liquid will mainly dictate the time required for drying, which can range from a few hours to several days (e.g. 40-60hrs).
  • a secondary drying stage may be carried out at about 0-40° C, depending primarily on the type and size of container and the type of protein employed. However, it was found herein that a secondary drying step may not be necessary.
  • the shelf temperature throughout the entire water removal phase of lyophilization may be from about 15-30° C. (e.g., about 20° C).
  • the time and pressure required for secondary drying will be that which produces a suitable lyophilized cake, dependent, e.g., on the temperature and other parameters.
  • the secondary drying time is dictated by the desired residual moisture level in the product and typically takes at least about 5 hours (e.g. 10-15 hours).
  • the pressure may be the same as that employed during the primary drying step. Freeze- drying conditions can be varied depending on the formulation and vial size.
  • the container in this instance may, for example, be a 3, 5, 10, 20, 50 or 100 cc vial.
  • lyophilization will result in a lyophilized formulation in which the moisture content thereof is less than about 5%, and preferably less than about 3%.
  • the lyophilized formulation may be reconstituted with a diluent such that the protein concentration in the reconstituted formulation is at least 50 mg/mL, for example from about 50 mg/mL to about 400 mg/mL, more preferably from about 80 mg/mL to about 300 mg/mL, and most preferably from about 90 mg/mL to about 150 mg/mL.
  • a diluent such that the protein concentration in the reconstituted formulation is at least 50 mg/mL, for example from about 50 mg/mL to about 400 mg/mL, more preferably from about 80 mg/mL to about 300 mg/mL, and most preferably from about 90 mg/mL to about 150 mg/mL.
  • Such high protein concentrations in the reconstituted formulation are considered to be particularly useful where subcutaneous delivery of the reconstituted formulation is intended.
  • the protein concentration in the reconstituted formulation is significantly higher than that in the pre- lyophilized formulation.
  • the protein concentration in the reconstituted formulation may be about 2-40 times, preferably 3-10 times and most preferably 3-6 times (e.g. at least three fold or at least four fold) that of the pre- lyophilized formulation.
  • the disclosure also provides combination pharmaceutical compositions suitable for monotherapy or combination therapy that comprises substantially homogeneous glycoantibodies described herein and other antibodies and/or other therapeutic agents.
  • the pharmaceutical composition can be administered as coformulation or used in co-administration therapeutic regimen.
  • Reconstitution generally takes place at a temperature of about 25° C. to ensure complete hydration, although other temperatures may be employed as desired.
  • the time required for reconstitution will depend, e.g., on the type of diluent, amount of excipient(s) and protein.
  • Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffered solution (e.g. phosphate-buffered saline), sterile saline solution, Ringer's solution or dextrose solution.
  • BWFI bacteriostatic water for injection
  • pH buffered solution e.g. phosphate-buffered saline
  • sterile saline solution e.g. phosphate-buffered saline
  • Ringer's solution or dextrose solution e.g. sterile saline
  • the diluent optionally contains a preservative. Exemplary preservatives have been
  • the amount of preservative employed is determined by assessing different preservative concentrations for compatibility with the protein and preservative efficacy testing.
  • the preservative is an aromatic alcohol (such as benzyl alcohol)
  • it can be present in an amount from about 0.1-2.0% and preferably from about 0.5-1.5%, but most preferably about 1.0-1.2%.
  • the reconstituted formulation has less than 6000 particles per vial which are >10 ⁇ m size.
  • the invention provides immunoconjugates, or antibody-drug conjugates (ADC), comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, a drug, a growth inhibitory agent, a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate).
  • a cytotoxic agent such as a chemotherapeutic agent, a drug, a growth inhibitory agent, a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate).
  • Toxins used in antibody -toxin conjugates include bacterial toxins such as diphtheria toxin, plant toxins such as ricin, small molecule toxins such as geldanamycin (Mandler et al (2000) Jour, of the Nat. Cancer Inst. 92(19): 1573-1581 ; Mandler et al (2000) Bioorganic & Med. Chem. Letters 10: 1025-1028; Mandler et al (2002) Bioconjugate Chem. 13 :786-791), maytansinoids (EP 1391213; Liu et al, (1996) Proc. Natl. Acad. Sci.
  • cytotoxic and cytostatic effects may affect their cytotoxic and cytostatic effects by mechanisms including tubutin binding, DNA binding, or topoisomerase inhibition. Some cytotoxic drugs tend to be inactive or less active when conjugated to large antibodies or protein receptor ligands.
  • Antibodies of the invention can be further modified to contain additional nonproteinaceous moieties that are known in the art and readily available.
  • the moieties suitable for derivatization of the antibody are water soluble polymers.
  • water soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-l,3-dioxolane, poly-l,3,6-trioxane, ethylene/maleic anhydride copolymer, polyaminoacids (either homopolymers or random copolymers), and dextran or poly(n- vinyl pyrrolidone)polyethylene glycol, propropylene glycol homopolymers, prolypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols (e.g., gly
  • Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water.
  • the polymer may be of any molecular weight, and may be branched or unbranched.
  • the number of polymers attached to the antibody may vary, and if more than one polymer is attached, the polymers can be the same or different molecules. In general, the number and/or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used in a therapy under defined conditions, etc.
  • conjugates of an antibody and nonproteinaceous moiety that may be selectively heated by exposure to radiation are provided.
  • the nonproteinaceous moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. 102: 1 1600- 11605 (2005)).
  • the radiation may be of any wavelength, and includes, but is not limited to, wavelengths that do not harm ordinary cells, but which heat the nonproteinaceous moiety to a temperature at which cells proximal to the antibody-nonproteinaceous moiety are killed.
  • rheumatoid arthritis is an autoimmune disease affecting more than two million people in the United States. RA causes chronic inflammation of the joints and typically is a progressive illness that has the potential to cause joint destruction and functional disability. The cause of rheumatoid arthritis is unknown, although genetic predisposition, infectious agents and environmental factors have all been implicated in the etiology of the disease. In active RA, symptoms can include fatigue, lack of appetite, low grade fever, muscle and joint aches and stiffness. Also during disease flare ups, joints frequently become red, swollen, painful and tender, due to inflammation of the synovium. Furthermore, since RA is a systemic disease,
  • inflammation can affect organs and areas of the body other than the joints, including glands of the eyes and mouth, the lung lining, the pericardium, and blood vessels.
  • Tumor necrosis factor alpha is a cytokine produced by numerous cell types, including monocytes and macrophages, that was originally identified based on its ability to induce the necrosis of certain mouse tumors. Subsequently, a factor termed cachectin, associated with cachexia, was shown to be identical to TNF-a. TNF-a has been implicated in the pathophysiology of a variety of other human diseases and disorders, including shock, sepsis, infections, autoimmune diseases, RA, Crohn's disease, transplant rejection and graft-versus-host disease.
  • the present disclosure provides a method of treating a TNF-mediated inflammatory disease in a human, the method comprising administering an inflammatory effective amount of an exemplary anti-TNFa glycoantibody or antigen binding fragment thereof and the
  • the exemplary inflammatory disease accordingly to the method of the invention can be selected from the group of inflammatory bowel disease (including ulcerative colitis and Crohn's disease), rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, and juvenile idiopathic arthritis.
  • inflammatory bowel disease including ulcerative colitis and Crohn's disease
  • rheumatoid arthritis including ulcerative colitis and Crohn's disease
  • psoriasis psoriatic arthritis
  • ankylosing spondylitis and juvenile idiopathic arthritis.
  • Adalimumab(Humira®) purchased from Abbvie
  • DTT dithiothreitol
  • IAA Iodoacetamide
  • microcentrifugation with spin column (lOkDa protein MW cut-off).
  • the trypsin digestion was performed for 4 hours at 37 °C using an enzyme: protein ratio of 1 : 25 (w/w).
  • Sample was frozen at -20 °C for LC-MS/MS analysis.
  • glycopeptide quantification by m/z 204 oxonium ion (HexNAc) monitoring was performed using a 4000 QTrap triple quadrupole mass spectrometer (AB Sciex) with Aglient 1200 HPLC system.
  • Adalimumab (2.5 mg) in a sodium phosphate buffer (50 mM, pH 7.0, 1.25 mL) was incubated with Endo S (125 ⁇ g) at 37°C for 5 h to yield a disaccharide (GlcNAc-Fuc) on Fc of Adalimumab (anti-TNFa GAb200).
  • Endo S 125 ⁇ g
  • Endo S 125 ⁇ g
  • Adalimumab anti-TNFa GAb200
  • EEQYNSTYR were analyzed using nanospray LC/MS to verify the glycosylation pattern of GAb200. Result of N-glycan profiling of anti-TNFa GAb200 is shown in Figure 1.
  • Adalimumab (2.5 mg) in a sodium phosphate buffer (50 mM, pH 7.0, 1.25 mL) was incubated with a mixture of Endo S (125 ⁇ g) and Bacteroides alpha-L-fucosidase (2.5 mg) at 37°C for 5 h to yield a monosaccharide (GlcNAc) on Fc of Adalimumab (anti-TNFa GAb201).
  • the reaction mixture was subject to affinity chromatography on a column of protein A-agarose resin (1 mL) that was pre-equilibrated with a sodium phosphate buffer (20 mM, pH 7.0).
  • the column was washed with a sodium phosphate buffer (20 mM, pH 7.0, 10 mL).
  • the bound IgG was released with glycine-HCl (50 mM, pH 3.0, 10 mL), and the elution fractions were immediately neutralized with Tris-Cl buffer (1.0 M, pH 8.3).
  • the fractions containing the Fc fragments were combined and concentrated by centrifugal filtration (Amicon Ultra centrifugal filter, Millipore, Billerica, MA) to give anti-TNFa GAb201.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb201.
  • Glycan oxazoline-101 was added to a mixture of endoglycosidase and GAb 201 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAblOl.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAblOl.
  • Glycan oxazoline-104 was added to a mixture of endoglycosidase and GAb 201 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb 104.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAM04.
  • Glycan oxazoline-107 was added to a mixture of endoglycosidase and GAb 201 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb 107.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb 107.
  • Glycan oxazoline-109 was added to a mixture of endoglycosidase and GAb 201 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb 109.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb 109.
  • sialylglycopeptide (SGP) was according to the published method. Briefly, the phenol extraction of hen's egg yolk was centrifuged, filtrated, and purified by the chromatographic columns, including Sephadex G-50, Sephadex G-25, DEAE- Toyoperarl 650M, CM-Sephadex C-25 and Sephadex G-25. A solution of sialylglycopeptide (SGP) (52 mg) in a sodium phosphate buffer (50 mM, pH 6.0, 5mM) was incubated with the Endo M (53 ⁇ g) at 37 °C.
  • SGP sodium phosphate buffer
  • Glycan oxazoline-401 was added to a mixture of endoglycosidase and GAb 200 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb401.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb401. Results of N-glycan profiling is shown in Figure 1.
  • Glycan oxazoline-404 was added to a mixture of endoglycosidase and GAb 200 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb404.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb404.
  • Glycan oxazoline-407 was added to a mixture of endoglycosidase and GAb 200 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb407.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb407.
  • Glycan oxazoline-409 was added to a mixture of endoglycosidase and GAb 200 in 50 mM Tris buffer (pH 7.8) and incubated for an hour at room temperature.
  • the reaction mixture was purified with protein A affinity column, followed by amanion exchange column capto Q to collect the desired product, anti-TNFa GAb409.
  • the product was trypsinized, and the glycopeptides, TKPREEQYNSTYR and EEQYNSTYR, were analyzed using nanospray LC/MS to confirm the glycosylation pattern of GAb409.
  • the FcyRIIIA recombinant protein was purified from transfected HEK-293 cell line, and then prepared at 0.5 ug/mL in ELISA coating buffer (50 mM Na 2 C0 3 , 50 mM NaHC0 3 , pHIO).
  • Anti-TNFa GAbs were 5-fold titrated from 150 nM to 1.54* 10 "5 nM in 2% BSA/PBST, and then applied to the ELISA plates in which recombinant FcyRIIIA were pre-immobilized. After 1 hour of incubation at RT, the plates were treated with anti-human IgG-HRP in 2% BSA/TBST at RT for 0.5hr.
  • Fc receptor binding in exemplary embodiments is increased by at least about 1.35-fold, 1.5 fold, 1.75 fold, 2-fold, 2.5 fold, 3-fold or higher.

Abstract

La présente invention concerne une nouvelle classe d'anticorps monoclonaux anti-TNFα ou des fragments de liaison de l'antigène comprenant une population homogène de molécules IgG anti-TNFα ayant le même N-glycane sur chaque Fc. Les anticorps de l'invention peut être produits à partir d'anticorps monoclonaux anti-TNFα par glycomodification de Fc. Les glycoanticorps de l'invention peuvent avoir des valeurs thérapeutique améliorées par rapport à celles d'anticorps monoclonaux correspondants qui n'ont pas été glycomodifiés.
PCT/US2015/032737 2014-05-28 2015-05-27 Glycoanticorps anti-thf-alpha et leurs utilisations WO2015184001A1 (fr)

Priority Applications (7)

Application Number Priority Date Filing Date Title
CN201580027936.6A CN106714829A (zh) 2014-05-28 2015-05-27 抗TNF‑α醣抗体及其用途
JP2016569732A JP7063538B2 (ja) 2014-05-28 2015-05-27 抗ＴＮＦα糖操作抗体群およびその使用
CA2950433A CA2950433A1 (fr) 2014-05-28 2015-05-27 Glycoanticorps anti-thf-alpha et leurs utilisations
EP15800059.6A EP3154582A4 (fr) 2014-05-28 2015-05-27 Glycoanticorps anti-thf-alpha et leurs utilisations
KR1020167036499A KR102494193B1 (ko) 2014-05-28 2015-05-27 항-ｔｎｆ-알파 글리코항체 및 이의 용도
AU2015267044A AU2015267044A1 (en) 2014-05-28 2015-05-27 Anti-TNF-alpha glycoantibodies and uses thereof
IL249184A IL249184B (en) 2014-05-28 2016-11-24 Anti-TNF-alpha glycoantibodies and their uses

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US201462003908P 2014-05-28 2014-05-28
US62/003,908 2014-05-28
US201462020199P 2014-07-02 2014-07-02
US62/020,199 2014-07-02
US201562110338P 2015-01-30 2015-01-30
US62/110,338 2015-01-30

Publications (2)

Publication Number Publication Date
WO2015184001A1 true WO2015184001A1 (fr) 2015-12-03
WO2015184001A8 WO2015184001A8 (fr) 2021-10-21

Family

ID=54699735

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2015/032737 WO2015184001A1 (fr) 2014-05-28 2015-05-27 Glycoanticorps anti-thf-alpha et leurs utilisations

Country Status (10)

Country Link
US (1) US11332523B2 (fr)
EP (1) EP3154582A4 (fr)
JP (2) JP7063538B2 (fr)
KR (1) KR102494193B1 (fr)
CN (1) CN106714829A (fr)
AU (1) AU2015267044A1 (fr)
CA (1) CA2950433A1 (fr)
IL (1) IL249184B (fr)
TW (1) TWI732738B (fr)
WO (1) WO2015184001A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3257866A1 (fr) * 2016-06-17 2017-12-20 Academisch Medisch Centrum Anticorps anti-tnf modifiés et leur utilisation dans le traitement de la maladie intestinale inflammatoire
WO2019234021A1 (fr) * 2018-06-04 2019-12-12 Limmatech Biologics Ag Anticorps monoclonal glycomodifié

Families Citing this family (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7960139B2 (en) 2007-03-23 2011-06-14 Academia Sinica Alkynyl sugar analogs for the labeling and visualization of glycoconjugates in cells
DK2318832T3 (da) 2008-07-15 2014-01-20 Academia Sinica Glycan-arrays på PTFE-lignende aluminiumcoatede objektglas og relaterede fremgangsmåder
US11377485B2 (en) * 2009-12-02 2022-07-05 Academia Sinica Methods for modifying human antibodies by glycan engineering
US10087236B2 (en) * 2009-12-02 2018-10-02 Academia Sinica Methods for modifying human antibodies by glycan engineering
WO2011130332A1 (fr) 2010-04-12 2011-10-20 Academia Sinica Puces au glycane pour la recherche par criblage haut débit de virus
US10130714B2 (en) 2012-04-14 2018-11-20 Academia Sinica Enhanced anti-influenza agents conjugated with anti-inflammatory activity
EP2885311B1 (fr) 2012-08-18 2020-01-01 Academia Sinica Sondes perméables aux cellules pour l'identification et l'imagerie de sialidases
US10086054B2 (en) 2013-06-26 2018-10-02 Academia Sinica RM2 antigens and use thereof
EP3013347B1 (fr) 2013-06-27 2019-12-11 Academia Sinica Conjugués de glycane et leur utilisation
CN105682666B (zh) 2013-09-06 2021-06-01 中央研究院 使用醣脂激活人类iNKT细胞
EP3094352B1 (fr) 2014-01-16 2020-09-23 Academia Sinica Compositions et méthodes pour traiter et détecter des cancers
US10150818B2 (en) 2014-01-16 2018-12-11 Academia Sinica Compositions and methods for treatment and detection of cancers
TWI687428B (zh) 2014-03-27 2020-03-11 中央研究院 反應性標記化合物及其用途
US10005847B2 (en) 2014-05-27 2018-06-26 Academia Sinica Anti-HER2 glycoantibodies and uses thereof
AU2015267047A1 (en) 2014-05-27 2017-01-05 Academia Sinica Anti-CD20 glycoantibodies and uses thereof
US10118969B2 (en) 2014-05-27 2018-11-06 Academia Sinica Compositions and methods relating to universal glycoforms for enhanced antibody efficacy
EP3904388A1 (fr) * 2014-05-27 2021-11-03 Academia Sinica Fucosidase issue de bacteroïdes et ses procédés d'utilisation
WO2015184001A1 (fr) 2014-05-28 2015-12-03 Academia Sinica Glycoanticorps anti-thf-alpha et leurs utilisations
KR102422375B1 (ko) 2014-09-08 2022-07-18 아카데미아 시니카 당지질을 사용한 인간 iNKT 세포 활성화
US10495645B2 (en) 2015-01-16 2019-12-03 Academia Sinica Cancer markers and methods of use thereof
US9975965B2 (en) 2015-01-16 2018-05-22 Academia Sinica Compositions and methods for treatment and detection of cancers
AU2015378564A1 (en) 2015-01-24 2017-07-13 Academia Sinica Novel glycan conjugates and methods of use thereof
WO2017156192A1 (fr) 2016-03-08 2017-09-14 Academia Sinica Procédés de synthèse modulaire de n-glycanes et de puces à n-glycanes
JP7213549B2 (ja) 2016-08-22 2023-01-27 シーエイチオー ファーマ インコーポレイテッド 抗体、結合性断片、および使用の方法
CN116903738A (zh) * 2022-08-02 2023-10-20 北京绿竹生物技术股份有限公司 一种低甘露糖型抗人肿瘤坏死因子-α单抗及其用途

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6528286B1 (en) * 1998-05-29 2003-03-04 Genentech, Inc. Mammalian cell culture process for producing glycoproteins
US20060073161A1 (en) * 1999-04-09 2006-04-06 Breton Gary L Nucleic acid and amino acid sequences relating to Bacteroides fragilis for diagnostics and therapeutics
US20100173323A1 (en) * 2006-06-09 2010-07-08 University Of Maryland, Baltimore Glycosylation engineered antibody therapy
US20110263828A1 (en) * 2009-12-02 2011-10-27 Academia Sinica Methods for modifying human antibodies by glycan engineering
US20120226024A1 (en) * 2011-03-03 2012-09-06 Lai-Xi Wang Core fucosylated glycopeptides and glycoproteins: chemoenzymatic synthesis and uses thereof
US20130230886A1 (en) * 2007-12-31 2013-09-05 Christian Votsmeier Antibodies to TNF Alpha
WO2013181585A2 (fr) * 2012-06-01 2013-12-05 Momenta Pharmaceuticals, Inc. Méthodes associées à l'adalimumab
US20130337018A1 (en) * 2007-10-02 2013-12-19 Avaxia Biologics, Incorporated Antibody therapy for use in the digestive tract

Family Cites Families (370)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US3896111A (en) 1973-02-20 1975-07-22 Research Corp Ansa macrolides
US4151042A (en) 1977-03-31 1979-04-24 Takeda Chemical Industries, Ltd. Method for producing maytansinol and its derivatives
US4137230A (en) 1977-11-14 1979-01-30 Takeda Chemical Industries, Ltd. Method for the production of maytansinoids
USRE30985E (en) 1978-01-01 1982-06-29 Serum-free cell culture media
US4307016A (en) 1978-03-24 1981-12-22 Takeda Chemical Industries, Ltd. Demethyl maytansinoids
US4265814A (en) 1978-03-24 1981-05-05 Takeda Chemical Industries Matansinol 3-n-hexadecanoate
JPS5562090A (en) 1978-10-27 1980-05-10 Takeda Chem Ind Ltd Novel maytansinoid compound and its preparation
US4256746A (en) 1978-11-14 1981-03-17 Takeda Chemical Industries Dechloromaytansinoids, their pharmaceutical compositions and method of use
JPS55164687A (en) 1979-06-11 1980-12-22 Takeda Chem Ind Ltd Novel maytansinoid compound and its preparation
JPS5566585A (en) 1978-11-14 1980-05-20 Takeda Chem Ind Ltd Novel maytansinoid compound and its preparation
JPS55102583A (en) 1979-01-31 1980-08-05 Takeda Chem Ind Ltd 20-acyloxy-20-demethylmaytansinoid compound
JPS55162791A (en) 1979-06-05 1980-12-18 Takeda Chem Ind Ltd Antibiotic c-15003pnd and its preparation
JPS55164685A (en) 1979-06-08 1980-12-22 Takeda Chem Ind Ltd Novel maytansinoid compound and its preparation
JPS55164686A (en) 1979-06-11 1980-12-22 Takeda Chem Ind Ltd Novel maytansinoid compound and its preparation
US4309428A (en) 1979-07-30 1982-01-05 Takeda Chemical Industries, Ltd. Maytansinoids
JPS5645483A (en) 1979-09-19 1981-04-25 Takeda Chem Ind Ltd C-15003phm and its preparation
JPS5645485A (en) 1979-09-21 1981-04-25 Takeda Chem Ind Ltd Production of c-15003pnd
EP0028683A1 (fr) 1979-09-21 1981-05-20 Takeda Chemical Industries, Ltd. Antibiotique C-15003 PHO et sa préparation
US4270537A (en) 1979-11-19 1981-06-02 Romaine Richard A Automatic hypodermic syringe
US4376110A (en) 1980-08-04 1983-03-08 Hybritech, Incorporated Immunometric assays using monoclonal antibodies
WO1982001188A1 (fr) 1980-10-08 1982-04-15 Takeda Chemical Industries Ltd Composes 4,5-deoxymaytansinoide et leur procede de preparation
US4450254A (en) 1980-11-03 1984-05-22 Standard Oil Company Impact improvement of high nitrile resins
US4419446A (en) 1980-12-31 1983-12-06 The United States Of America As Represented By The Department Of Health And Human Services Recombinant DNA process utilizing a papilloma virus DNA as a vector
US4315929A (en) 1981-01-27 1982-02-16 The United States Of America As Represented By The Secretary Of Agriculture Method of controlling the European corn borer with trewiasine
US4313946A (en) 1981-01-27 1982-02-02 The United States Of America As Represented By The Secretary Of Agriculture Chemotherapeutically active maytansinoids from Trewia nudiflora
JPS57192389A (en) 1981-05-20 1982-11-26 Takeda Chem Ind Ltd Novel maytansinoid
US4596792A (en) 1981-09-04 1986-06-24 The Regents Of The University Of California Safe vaccine for hepatitis containing polymerized serum albumin
US4741900A (en) 1982-11-16 1988-05-03 Cytogen Corporation Antibody-metal ion complexes
US4601978A (en) 1982-11-24 1986-07-22 The Regents Of The University Of California Mammalian metallothionein promoter system
US4560655A (en) 1982-12-16 1985-12-24 Immunex Corporation Serum-free cell culture medium and process for making same
US4657866A (en) 1982-12-21 1987-04-14 Sudhir Kumar Serum-free, synthetic, completely chemically defined tissue culture media
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4767704A (en) 1983-10-07 1988-08-30 Columbia University In The City Of New York Protein-free culture medium
US4599230A (en) 1984-03-09 1986-07-08 Scripps Clinic And Research Foundation Synthetic hepatitis B virus vaccine including both T cell and B cell determinants
US4599231A (en) 1984-03-09 1986-07-08 Scripps Clinic And Research Foundation Synthetic hepatitis B virus vaccine including both T cell and B cell determinants
US4965199A (en) 1984-04-20 1990-10-23 Genentech, Inc. Preparation of functional human factor VIII in mammalian cells using methotrexate based selection
US4970198A (en) 1985-10-17 1990-11-13 American Cyanamid Company Antitumor antibiotics (LL-E33288 complex)
US4596556A (en) 1985-03-25 1986-06-24 Bioject, Inc. Hypodermic injection apparatus
US4601903A (en) 1985-05-01 1986-07-22 The United States Of America As Represented By The Department Of Health And Human Services Vaccine against Neisseria meningitidis Group B serotype 2 invasive disease
GB8516415D0 (en) 1985-06-28 1985-07-31 Celltech Ltd Culture of animal cells
US4676980A (en) 1985-09-23 1987-06-30 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Target specific cross-linked heteroantibodies
US4927762A (en) 1986-04-01 1990-05-22 Cell Enterprises, Inc. Cell culture medium with antioxidant
US5567610A (en) 1986-09-04 1996-10-22 Bioinvent International Ab Method of producing human monoclonal antibodies and kit therefor
US5811128A (en) 1986-10-24 1998-09-22 Southern Research Institute Method for oral or rectal delivery of microencapsulated vaccines and compositions therefor
US5075109A (en) 1986-10-24 1991-12-24 Southern Research Institute Method of potentiating an immune response
US4886499A (en) 1986-12-18 1989-12-12 Hoffmann-La Roche Inc. Portable injection appliance
IL85035A0 (en) 1987-01-08 1988-06-30 Int Genetic Eng Polynucleotide molecule,a chimeric antibody with specificity for human b cell surface antigen,a process for the preparation and methods utilizing the same
US5079233A (en) 1987-01-30 1992-01-07 American Cyanamid Company N-acyl derivatives of the LL-E33288 antitumor antibiotics, composition and methods for using the same
GB8704027D0 (en) 1987-02-20 1987-03-25 Owen Mumford Ltd Syringe needle combination
DE3883899T3 (de) 1987-03-18 1999-04-22 Sb2 Inc Geänderte antikörper.
US4849222A (en) 1987-03-24 1989-07-18 The Procter & Gamble Company Mixtures for treating hypercholesterolemia
US4790824A (en) 1987-06-19 1988-12-13 Bioject, Inc. Non-invasive hypodermic injection device
US4940460A (en) 1987-06-19 1990-07-10 Bioject, Inc. Patient-fillable and non-invasive hypodermic injection device assembly
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
US4975278A (en) 1988-02-26 1990-12-04 Bristol-Myers Company Antibody-enzyme conjugates in combination with prodrugs for the delivery of cytotoxic agents to tumor cells
US5004697A (en) 1987-08-17 1991-04-02 Univ. Of Ca Cationized antibodies for delivery through the blood-brain barrier
US5770701A (en) 1987-10-30 1998-06-23 American Cyanamid Company Process for preparing targeted forms of methyltrithio antitumor agents
US5606040A (en) 1987-10-30 1997-02-25 American Cyanamid Company Antitumor and antibacterial substituted disulfide derivatives prepared from compounds possessing a methyl-trithio group
US5053394A (en) 1988-09-21 1991-10-01 American Cyanamid Company Targeted forms of methyltrithio antitumor agents
JP2670680B2 (ja) 1988-02-24 1997-10-29 株式会社ビーエムジー 生理活性物質含有ポリ乳酸系微小球およびその製造法
US5339163A (en) 1988-03-16 1994-08-16 Canon Kabushiki Kaisha Automatic exposure control device using plural image plane detection areas
JPH01287029A (ja) 1988-05-13 1989-11-17 Mect Corp 新規抗ウィルス剤
ATE135397T1 (de) 1988-09-23 1996-03-15 Cetus Oncology Corp Zellenzuchtmedium für erhöhtes zellenwachstum, zur erhöhung der langlebigkeit und expression der produkte
GB8823869D0 (en) 1988-10-12 1988-11-16 Medical Res Council Production of antibodies
FR2638359A1 (fr) 1988-11-03 1990-05-04 Tino Dalto Guide de seringue avec reglage de la profondeur de penetration de l'aiguille dans la peau
US5175384A (en) 1988-12-05 1992-12-29 Genpharm International Transgenic mice depleted in mature t-cells and methods for making transgenic mice
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
DE3920358A1 (de) 1989-06-22 1991-01-17 Behringwerke Ag Bispezifische und oligospezifische, mono- und oligovalente antikoerperkonstrukte, ihre herstellung und verwendung
DE69029036T2 (de) 1989-06-29 1997-05-22 Medarex Inc Bispezifische reagenzien für die aids-therapie
US5690938A (en) 1989-07-07 1997-11-25 Oravax, Inc. Oral immunization with multiple particulate antigen delivery system
US5518725A (en) 1989-09-25 1996-05-21 University Of Utah Research Foundation Vaccine compositions and method for induction of mucosal immune response via systemic vaccination
US5208020A (en) 1989-10-25 1993-05-04 Immunogen Inc. Cytotoxic agents comprising maytansinoids and their therapeutic use
CA2026147C (fr) 1989-10-25 2006-02-07 Ravi J. Chari Agents cytotoxiques comprenant des maytansinoides et leur usage therapeutique
US5238843A (en) 1989-10-27 1993-08-24 Genencor International, Inc. Method for cleaning a surface on which is bound a glycoside-containing substance
US5064413A (en) 1989-11-09 1991-11-12 Bioject, Inc. Needleless hypodermic injection device
US5312335A (en) 1989-11-09 1994-05-17 Bioject Inc. Needleless hypodermic injection device
EP1690935A3 (fr) 1990-01-12 2008-07-30 Abgenix, Inc. Génération d'anticorps xenogéniques
US5061620A (en) 1990-03-30 1991-10-29 Systemix, Inc. Human hematopoietic stem cell
US5268164A (en) 1990-04-23 1993-12-07 Alkermes, Inc. Increasing blood-brain barrier permeability with permeabilizer peptides
US5112596A (en) 1990-04-23 1992-05-12 Alkermes, Inc. Method for increasing blood-brain barrier permeability by administering a bradykinin agonist of blood-brain barrier permeability
US5643577A (en) 1990-04-24 1997-07-01 The University Of Newcastle Research Associates Limited Oral vaccine comprising antigen surface-associated with red blood cells
AP249A (en) 1990-04-24 1993-03-17 Biota Scient Management Pty Ltd Anti-viral compounds.
US5229275A (en) 1990-04-26 1993-07-20 Akzo N.V. In-vitro method for producing antigen-specific human monoclonal antibodies
US5427908A (en) 1990-05-01 1995-06-27 Affymax Technologies N.V. Recombinant library screening methods
KR0149181B1 (ko) 1990-06-29 1998-08-17 데이비드 알, 맥지 형질전환된 미생물에 의한 멜라닌의 제조방법
US5190521A (en) 1990-08-22 1993-03-02 Tecnol Medical Products, Inc. Apparatus and method for raising a skin wheal and anesthetizing skin
US5625126A (en) 1990-08-29 1997-04-29 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5633425A (en) 1990-08-29 1997-05-27 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5661016A (en) 1990-08-29 1997-08-26 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
KR100272077B1 (ko) 1990-08-29 2000-11-15 젠팜인터내셔날,인코포레이티드 이종 항체를 생산할 수 있는 전이유전자를 가진 인간이외의 동물
US5545806A (en) 1990-08-29 1996-08-13 Genpharm International, Inc. Ransgenic non-human animals for producing heterologous antibodies
US5714374A (en) 1990-09-12 1998-02-03 Rutgers University Chimeric rhinoviruses
US5122469A (en) 1990-10-03 1992-06-16 Genentech, Inc. Method for culturing Chinese hamster ovary cells to improve production of recombinant proteins
WO1992006691A1 (fr) 1990-10-19 1992-04-30 Biota Scientific Management Pty. Ltd. Composes anti-viraux qui lient le site actif de la neuraminidase de la grippe et presentent une activite in vivo contre l'orthomixovirus et le paramyxovirus
US5264365A (en) 1990-11-09 1993-11-23 Board Of Regents, The University Of Texas System Protease-deficient bacterial strains for production of proteolytically sensitive polypeptides
US5508192A (en) 1990-11-09 1996-04-16 Board Of Regents, The University Of Texas System Bacterial host strains for producing proteolytically sensitive polypeptides
ATE164395T1 (de) 1990-12-03 1998-04-15 Genentech Inc Verfahren zur anreicherung von proteinvarianten mit geänderten bindungseigenschaften
US5527288A (en) 1990-12-13 1996-06-18 Elan Medical Technologies Limited Intradermal drug delivery device and method for intradermal delivery of drugs
US5571894A (en) 1991-02-05 1996-11-05 Ciba-Geigy Corporation Recombinant antibodies specific for a growth factor receptor
WO1992019971A1 (fr) 1991-04-30 1992-11-12 Alkermes, Inc. Anticorps cationises contre des proteines intracellulaires
DK0590058T3 (da) 1991-06-14 2004-03-29 Genentech Inc Humaniseret heregulin-antistof
GB9114948D0 (en) 1991-07-11 1991-08-28 Pfizer Ltd Process for preparing sertraline intermediates
GB9118204D0 (en) 1991-08-23 1991-10-09 Weston Terence E Needle-less injector
SE9102652D0 (sv) 1991-09-13 1991-09-13 Kabi Pharmacia Ab Injection needle arrangement
WO1993006217A1 (fr) 1991-09-19 1993-04-01 Genentech, Inc. EXPRESSION DANS L'E. COLI DE FRAGMENTS D'ANTICORPS POSSEDANT AU MOINS UNE CYSTEINE PRESENTE SOUS FORME D'UN THIOL LIBRE, ET LEUR UTILISATION DANS LA PRODUCTION D'ANTICORPS BIFONCTIONNELS F(ab')¿2?
ES2136092T3 (es) 1991-09-23 1999-11-16 Medical Res Council Procedimientos para la produccion de anticuerpos humanizados.
DK0605522T3 (da) 1991-09-23 2000-01-17 Medical Res Council Fremgangsmåde til fremstilling af humaniserede antistoffer
US5362852A (en) 1991-09-27 1994-11-08 Pfizer Inc. Modified peptide derivatives conjugated at 2-hydroxyethylamine moieties
US5587458A (en) 1991-10-07 1996-12-24 Aronex Pharmaceuticals, Inc. Anti-erbB-2 antibodies, combinations thereof, and therapeutic and diagnostic uses thereof
US5288502A (en) 1991-10-16 1994-02-22 The University Of Texas System Preparation and uses of multi-phase microspheres
WO1993008829A1 (fr) 1991-11-04 1993-05-13 The Regents Of The University Of California Compositions induisant la destruction de cellules infectees par l'hiv
WO1993009764A1 (fr) 1991-11-19 1993-05-27 Center For Innovative Technology Traitement des rhumes a l'aide d'une therapie associant des agents antimediatiques et virostatiques (covam)
JPH0826057B2 (ja) 1992-01-16 1996-03-13 株式会社ディ・ディ・エス研究所 シアル酸オリゴ糖誘導体及び微粒子キャリヤー
US5667988A (en) 1992-01-27 1997-09-16 The Scripps Research Institute Methods for producing antibody libraries using universal or randomized immunoglobulin light chains
EP1997894B1 (fr) 1992-02-06 2011-03-30 Novartis Vaccines and Diagnostics, Inc. Protéine de liaison biosynthétique pour un marqueur du cancer
US5328483A (en) 1992-02-27 1994-07-12 Jacoby Richard M Intradermal injection device with medication and needle guard
US5733743A (en) 1992-03-24 1998-03-31 Cambridge Antibody Technology Limited Methods for producing members of specific binding pairs
US5326856A (en) 1992-04-09 1994-07-05 Cytogen Corporation Bifunctional isothiocyanate derived thiocarbonyls as ligands for metal binding
ZA932522B (en) 1992-04-10 1993-12-20 Res Dev Foundation Immunotoxins directed against c-erbB-2(HER/neu) related surface antigens
JP2904647B2 (ja) 1992-06-12 1999-06-14 株式会社蛋白工学研究所 ５−ブロム−４−クロロインド−３−イル−２−シアル酸の製造方法
ATE348175T1 (de) 1992-07-17 2007-01-15 Dana Farber Cancer Inst Inc Method eder intrazellularen bindung von zielgerichteten molekülen
US5383851A (en) 1992-07-24 1995-01-24 Bioject Inc. Needleless hypodermic injection device
WO1994002178A1 (fr) 1992-07-27 1994-02-03 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Ciblage de liposomes sur la barriere hemato-encephalique
AU668423B2 (en) 1992-08-17 1996-05-02 Genentech Inc. Bispecific immunoadhesins
US5569189A (en) 1992-09-28 1996-10-29 Equidyne Systems, Inc. hypodermic jet injector
DE69328435T2 (de) 1992-10-22 2000-09-14 Kirin Brewery Neues sphingolglycolipid und verwendung davon
US5807722A (en) 1992-10-30 1998-09-15 Bioengineering Resources, Inc. Biological production of acetic acid from waste gases with Clostridium ljungdahlii
US5334144A (en) 1992-10-30 1994-08-02 Becton, Dickinson And Company Single use disposable needleless injector
US5736137A (en) 1992-11-13 1998-04-07 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma
WO1994011026A2 (fr) 1992-11-13 1994-05-26 Idec Pharmaceuticals Corporation Application therapeutique d'anticorps chimeriques et radio-marques contre l'antigene a differentiation restreinte des lymphocytes b humains pour le traitement du lymphome des cellules b
US5635483A (en) 1992-12-03 1997-06-03 Arizona Board Of Regents Acting On Behalf Of Arizona State University Tumor inhibiting tetrapeptide bearing modified phenethyl amides
JP3523285B2 (ja) 1993-01-22 2004-04-26 雪印乳業株式会社 糖分解酵素の製造法
US5780588A (en) 1993-01-26 1998-07-14 Arizona Board Of Regents Elucidation and synthesis of selected pentapeptides
US5374541A (en) 1993-05-04 1994-12-20 The Scripps Research Institute Combined use of β-galactosidase and sialyltransferase coupled with in situ regeneration of CMP-sialic acid for one pot synthesis of oligosaccharides
US20020037517A1 (en) 1993-05-28 2002-03-28 Hutchens T. William Methods for sequencing biopolymers
CA2163345A1 (fr) 1993-06-16 1994-12-22 Susan Adrienne Morgan Anticorps
WO1995011010A1 (fr) 1993-10-22 1995-04-27 Genentech, Inc. Compositions et procedes de microencapsulation d'antigenes a utiliser comme vaccins
US5369017A (en) 1994-02-04 1994-11-29 The Scripps Research Institute Process for solid phase glycopeptide synthesis
CA2181237A1 (fr) 1994-02-25 1995-08-31 Subramaniam Sabesan Acides sialiques 4-n-substitues et leurs sialosides
WO1995024176A1 (fr) 1994-03-07 1995-09-14 Bioject, Inc. Dispositif de remplissage d'ampoule
US5466220A (en) 1994-03-08 1995-11-14 Bioject, Inc. Drug vial mixing and transfer device
US5773001A (en) 1994-06-03 1998-06-30 American Cyanamid Company Conjugates of methyltrithio antitumor agents and intermediates for their synthesis
US5622701A (en) 1994-06-14 1997-04-22 Protein Design Labs, Inc. Cross-reacting monoclonal antibodies specific for E- and P-selectin
ATE306930T1 (de) 1994-08-12 2005-11-15 Immunomedics Inc Für b-zell-lymphom und leukämiezellen spezifische immunkonjugate und humane antikörper
US5639635A (en) 1994-11-03 1997-06-17 Genentech, Inc. Process for bacterial production of polypeptides
EP0794792A1 (fr) 1994-12-02 1997-09-17 Chiron Corporation Methode permettant d'induire une reponse immunitaire avec un anticorps bispecifique
US5663149A (en) 1994-12-13 1997-09-02 Arizona Board Of Regents Acting On Behalf Of Arizona State University Human cancer inhibitory pentapeptide heterocyclic and halophenyl amides
US5599302A (en) 1995-01-09 1997-02-04 Medi-Ject Corporation Medical injection system and method, gas spring thereof and launching device using gas spring
US5840523A (en) 1995-03-01 1998-11-24 Genetech, Inc. Methods and compositions for secretion of heterologous polypeptides
US5731168A (en) 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US6673533B1 (en) 1995-03-10 2004-01-06 Meso Scale Technologies, Llc. Multi-array multi-specific electrochemiluminescence testing
US5641870A (en) 1995-04-20 1997-06-24 Genentech, Inc. Low pH hydrophobic interaction chromatography for antibody purification
US5869046A (en) 1995-04-14 1999-02-09 Genentech, Inc. Altered polypeptides with increased half-life
WO1996036436A1 (fr) 1995-04-25 1996-11-21 Irori Matrices programmables a distance et dotees de memoires, et leur utilisation
DE69637481T2 (de) 1995-04-27 2009-04-09 Amgen Fremont Inc. Aus immunisierten Xenomäusen stammende menschliche Antikörper gegen IL-8
EP0823941A4 (fr) 1995-04-28 2001-09-19 Abgenix Inc Anticorps humains derives de xeno-souris immunisees
US5730723A (en) 1995-10-10 1998-03-24 Visionary Medical Products Corporation, Inc. Gas pressured needle-less injection device and method
US6265150B1 (en) 1995-06-07 2001-07-24 Becton Dickinson & Company Phage antibodies
US5714586A (en) 1995-06-07 1998-02-03 American Cyanamid Company Methods for the preparation of monomeric calicheamicin derivative/carrier conjugates
US5837234A (en) 1995-06-07 1998-11-17 Cytotherapeutics, Inc. Bioartificial organ containing cells encapsulated in a permselective polyether suflfone membrane
US5712374A (en) 1995-06-07 1998-01-27 American Cyanamid Company Method for the preparation of substantiallly monomeric calicheamicin derivative/carrier conjugates
JPH11510164A (ja) 1995-07-26 1999-09-07 マキシム ファーマシューティカルズ ポリヌクレオチドの粘膜送達
DE19544393A1 (de) 1995-11-15 1997-05-22 Hoechst Schering Agrevo Gmbh Synergistische herbizide Mischungen
US5893397A (en) 1996-01-12 1999-04-13 Bioject Inc. Medication vial/syringe liquid-transfer apparatus
WO1997038123A1 (fr) 1996-04-05 1997-10-16 Board Of Regents, The University Of Texas System Procedes de production de proteines eucaryotes, solubles, actives sur plan biologique et contenant des liaisons disulfure, a l'interieur de cellules bacteriennes
GB9607549D0 (en) 1996-04-11 1996-06-12 Weston Medical Ltd Spring-powered dispensing device
US5922845A (en) 1996-07-11 1999-07-13 Medarex, Inc. Therapeutic multispecific compounds comprised of anti-Fcα receptor antibodies
US6340702B1 (en) 1996-07-22 2002-01-22 Sankyo Company, Limited Neuraminic acid derivatives, their preparation and their medical use
US6506564B1 (en) 1996-07-29 2003-01-14 Nanosphere, Inc. Nanoparticles having oligonucleotides attached thereto and uses therefor
IL129762A0 (en) 1996-11-14 2000-02-29 Biota Scient Management Novel macromolecules methods for their preparation pharmaceutical compositions thereof and their use as anti-influenza agents
EP2305027B1 (fr) 1996-12-03 2014-07-02 Amgen Fremont Inc. Mammifères trangèniques obtenus par génie génétique contenant des loci des immunoglobulines humaines qui comprennent plusieurs régions de VH et de Vkappa, et anticorps aussi obtenus.
TW555562B (en) 1996-12-27 2003-10-01 Kirin Brewery Method for activation of human antigen-presenting cells, activated human antigen-presenting cells and use thereof
AU733282B2 (en) 1997-04-18 2001-05-10 Novartis Ag Neoglycoproteins
US5993412A (en) 1997-05-19 1999-11-30 Bioject, Inc. Injection apparatus
US6083715A (en) 1997-06-09 2000-07-04 Board Of Regents, The University Of Texas System Methods for producing heterologous disulfide bond-containing polypeptides in bacterial cells
JPH1135593A (ja) 1997-07-18 1999-02-09 Daikin Ind Ltd ２−フルオロフコシル−ｎ−アロイルグルコサミン誘導体及びその中間物、並びにそれらの製造方法
TW477783B (en) 1997-12-12 2002-03-01 Gilead Sciences Inc Novel compounds useful as neuraminidase inhibitors and pharmaceutical compositions containing same
IT1298087B1 (it) 1998-01-08 1999-12-20 Fiderm S R L Dispositivo per il controllo della profondita' di penetrazione di un ago, in particolare applicabile ad una siringa per iniezioni
JP2002507410A (ja) 1998-03-27 2002-03-12 プロルーム・リミテッド ルシフェラーゼ、蛍光タンパク質、ルシフェラーゼおよび蛍光タンパク質をコードする核酸および、診断、高処理スクリーニングおよび新規アイテムにおけるその使用
JP2002510481A (ja) 1998-04-02 2002-04-09 ジェネンテック・インコーポレーテッド 抗体変異体及びその断片
EP2261229A3 (fr) 1998-04-20 2011-03-23 GlycArt Biotechnology AG Modification par glycosylation d'anticorps aux fins d'amélioration de la cytotoxicité cellulaire dépendant des anticorps
US6455571B1 (en) 1998-04-23 2002-09-24 Abbott Laboratories Inhibitors of neuraminidases
JP4469933B2 (ja) 1998-05-06 2010-06-02 ジェネンテック， インコーポレイテッド イオン交換クロマトグラフィによるタンパク質精製
JP3773153B2 (ja) 1998-05-29 2006-05-10 独立行政法人理化学研究所 シアル酸誘導体
EP2306195A3 (fr) 1998-09-18 2012-04-25 Massachusetts Institute of Technology Applications biologiques de nano-cristaux semi-conducteur
FR2783523B1 (fr) 1998-09-21 2006-01-20 Goemar Lab Sa Fuco-oligosaccharides, enzyme pour leur preparation a partir des fucanes, bacterie productrice de l'enzyme et applications des fuco-oligosaccharides a la protection des plantes
US6994966B2 (en) 2000-02-17 2006-02-07 Glycominds Ltd. Combinatorial complex carbohydrate libraries and methods for the manufacture and uses thereof
US6696304B1 (en) 1999-02-24 2004-02-24 Luminex Corporation Particulate solid phase immobilized protein quantitation
AUPP913999A0 (en) 1999-03-12 1999-04-01 Biota Scientific Management Pty Ltd Novel chemical compounds and their use
US7854934B2 (en) 1999-08-20 2010-12-21 Sloan-Kettering Institute For Cancer Research Glycoconjugates, glycoamino acids, intermediates thereto, and uses thereof
US6824780B1 (en) 1999-10-29 2004-11-30 Genentech, Inc. Anti-tumor antibody compositions and methods of use
AUPQ422399A0 (en) 1999-11-24 1999-12-16 University Of New South Wales, The Method of screening transformed or transfected cells
US6727356B1 (en) 1999-12-08 2004-04-27 Epoch Pharmaceuticals, Inc. Fluorescent quenching detection reagents and methods
US7019129B1 (en) 2000-05-09 2006-03-28 Biosearch Technologies, Inc. Dark quenchers for donor-acceptor energy transfer
US7863020B2 (en) 2000-06-28 2011-01-04 Glycofi, Inc. Production of sialylated N-glycans in lower eukaryotes
US6514221B2 (en) 2000-07-27 2003-02-04 Brigham And Women's Hospital, Inc. Blood-brain barrier opening
US20020065259A1 (en) 2000-08-30 2002-05-30 Schatzberg Alan F. Glucocorticoid blocking agents for increasing blood-brain barrier permeability
AUPR001000A0 (en) 2000-09-08 2000-10-05 Biota Scientific Management Pty Ltd Novel chemical compounds and their use
US7034036B2 (en) 2000-10-30 2006-04-25 Pain Therapeutics, Inc. Inhibitors of ABC drug transporters at the blood-brain barrier
US20030083299A1 (en) 2000-11-04 2003-05-01 Ferguson Ian A. Non-invasive delivery of polypeptides through the blood-brain barrier
JP2002153272A (ja) 2000-11-24 2002-05-28 Inst Of Physical & Chemical Res 生体分子マイクロアレイ
US7754208B2 (en) 2001-01-17 2010-07-13 Trubion Pharmaceuticals, Inc. Binding domain-immunoglobulin fusion proteins
WO2002086096A2 (fr) 2001-01-23 2002-10-31 University Of Rochester Medical Center Procedes de production ou d'identification d'intracorps dans des cellules eucaryotes
US6884869B2 (en) 2001-04-30 2005-04-26 Seattle Genetics, Inc. Pentapeptide compounds and uses related thereto
DE10121982B4 (de) 2001-05-05 2008-01-24 Lts Lohmann Therapie-Systeme Ag Nanopartikel aus Protein mit gekoppeltem Apolipoprotein E zur Überwindung der Blut-Hirn-Schranke und Verfahren zu ihrer Herstellung
JP2002371087A (ja) 2001-06-15 2002-12-26 Mitsubishi Chemicals Corp 有機ホスホン酸
WO2003009812A2 (fr) 2001-07-25 2003-02-06 New York University Utilisation de glycosylceramides comme adjuvants pour des vaccins contre les infections et le cancer
ES2338218T3 (es) 2001-07-25 2010-05-05 Facet Biotech Corporation Formulacion farmacologica liofilizada estable de anticuerpos igg daclizumab.
ATE445838T1 (de) 2001-07-25 2009-10-15 Raptor Pharmaceutical Inc Zusammensetzungen und verfahren zur modulation des transports durch die blut-hirn-schranke
EP1423510A4 (fr) 2001-08-03 2005-06-01 Glycart Biotechnology Ag Variants de glycosylation d'anticorps presentant une cytotoxicite cellulaire accrue dependante des anticorps
BRPI0213207B1 (pt) 2001-10-10 2021-06-15 Novo Nordisk A/S Processo in vitro, isento de células, para a remodelagem de um peptídeo e processos para formar um conjugado entre peptídeos e um grupo de modificação
KR20040077655A (ko) 2001-10-19 2004-09-06 슈페리어 마이크로파우더스 엘엘씨 전자 형상 증착용 테잎 조성물
AUPR879601A0 (en) 2001-11-09 2001-12-06 Biota Scientific Management Pty Ltd Novel chemical compounds and their use
US20040018501A1 (en) 2001-11-21 2004-01-29 Keith Allen Methods and systems for analyzing complex biological systems
EP2295468B1 (fr) 2002-02-14 2015-07-08 Immunomedics, Inc. Anticorps anti-CD20 et protéines hybrides desdits anticorps, et méthodes d'utilisation
US20030162695A1 (en) 2002-02-27 2003-08-28 Schatzberg Alan F. Glucocorticoid blocking agents for increasing blood-brain barrier permeability
US7317091B2 (en) 2002-03-01 2008-01-08 Xencor, Inc. Optimized Fc variants
AU2003219277A1 (en) 2002-03-14 2003-09-29 Medical Research Council Intracellular antibodies
MXPA04011249A (es) 2002-05-14 2005-06-06 Chiron Srl Vacunas mucosales con adyuvante de quitosano y antigenos meningococicos.
WO2004005313A2 (fr) 2002-07-08 2004-01-15 Pliva - Istrazivacki Institut D.O.O. Nouveaux composes, compositions en tant que porteurs pour molecules actives anti-inflammatoires steroidiennes/non-steroidiennes, antineoplasiques et antivirales
US20080070324A1 (en) 2002-07-15 2008-03-20 Floyd Alton D Quantity control device for microscope slide staining assays
EP1391213A1 (fr) 2002-08-21 2004-02-25 Boehringer Ingelheim International GmbH Compositions et méthodes pour le traitement du cancer en utilisant un conjugué d'un anticorps contre le CD44 avec un maytansinoide et des agents chimiothérapeutiques
US20040062682A1 (en) 2002-09-30 2004-04-01 Rakow Neal Anthony Colorimetric sensor
LT3284753T (lt) 2002-10-17 2020-06-10 Genmab A/S Žmogaus monokloniniai antikūnai prieš cd20, skirti naudoti gydant išsėtinę sklerozę
AU2003302676A1 (en) 2002-12-03 2004-06-23 Blanchette Rockefeller Neurosciences Institute Artificial low-density lipoprotein carriers for transport of substances across the blood-brain barrier
PL212899B1 (pl) 2002-12-16 2012-12-31 Genentech Inc Humanizowane przeciwcialo, kompozycja zawierajaca to przeciwcialo, wyrób fabryczny, przeciwcialo lub jego fragment do zastosowania w sposobie indukowania apoptozy, izolowany kwas nukleinowy, wektor ekspresji, komórka gospodarza, sposób wytwarzania przeciwciala lub jego fragmentu, plynny preparat i zastosowanie przeciwciala do wytwarzania leku
AU2004204494B2 (en) 2003-01-09 2011-09-29 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
US8088387B2 (en) 2003-10-10 2012-01-03 Immunogen Inc. Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates, and methods of making said conjugates
AR044388A1 (es) 2003-05-20 2005-09-07 Applied Molecular Evolution Moleculas de union a cd20
US20040259142A1 (en) 2003-06-04 2004-12-23 Imperial College Innovations Limited Products and methods
EP2003196A3 (fr) 2003-06-09 2009-01-07 The Regents of the University of Michigan Compositions et procédés de diagnostic et de traitement des cancers
JP4148844B2 (ja) 2003-06-11 2008-09-10 ソニー・エリクソン・モバイルコミュニケーションズ株式会社 情報端末装置及び音声付画像ファイルの出力方法
WO2005025511A2 (fr) 2003-09-10 2005-03-24 Cedars-Sinai Medical Center Administration assistee par les canaux potassiques d'agents a travers la barriere hemato-encephalique
WO2005027990A2 (fr) 2003-09-15 2005-03-31 Univ Jefferson Biomateriaux ameliores et procede de fixation d'agents therapeutiques a ceux-ci
CA2539914A1 (fr) 2003-09-22 2005-04-07 Acidophil Llc Compositions a petites molecules et procedes destines a augmenter l'efficacite d'un medicament au moyen de ces compositions
WO2005033663A2 (fr) 2003-09-30 2005-04-14 Sequenom, Inc. Procedes de fabrication de substrats pour analyse par spectrometrie de masse et dispositifs associes
US20050221337A1 (en) 2003-10-02 2005-10-06 Massachusetts Institute Of Technology Microarrays and microspheres comprising oligosaccharides, complex carbohydrates or glycoproteins
HUE031632T2 (en) 2003-11-05 2017-07-28 Roche Glycart Ag Antigen binding molecules with enhanced Fc receptor binding affinity and effector function
BR122018071808B8 (pt) 2003-11-06 2020-06-30 Seattle Genetics Inc conjugado
US20050255491A1 (en) 2003-11-13 2005-11-17 Lee Frank D Small molecule and peptide arrays and uses thereof
EP1723422A2 (fr) 2004-03-05 2006-11-22 The Scripps Research Institute Jeux ordonnes de microechantillons de glycanes a haut rendement
US20050221397A1 (en) 2004-03-30 2005-10-06 Northern Advancement Center For Science & Technology RM2 antigen (beta1,4-GalNAc-disialyl-Lc4) as prostate cancer-associated antigen
EP1740946B1 (fr) 2004-04-20 2013-11-06 Genmab A/S Anticorps monoclonaux humains diriges contre cd20
ITMI20040928A1 (it) 2004-05-07 2004-08-07 Uni Di Bologna Dipartiment O D Procedura per la preparazione di coniugati della doxorubicina con l'albumina umana lattosaminata
JP2008504531A (ja) 2004-06-24 2008-02-14 ザ スクリップス リサーチ インスティテュート 切断可能なリンカーを有するアレイ
RU2361880C2 (ru) 2004-07-22 2009-07-20 Дженентек, Инк. Композиция антител к her2
US8022043B2 (en) 2004-08-27 2011-09-20 Albert Einstein College Of Medicine Of Yeshiva University Ceramide derivatives as modulators of immunity and autoimmunity
WO2006055925A2 (fr) 2004-11-19 2006-05-26 Swiss Federal Institute Of Technology Jeux ordonnes de microechantillons pour la detection d'analytes
WO2006064983A1 (fr) 2004-12-14 2006-06-22 Korea Research Institute Of Bioscience And Biotechnology Anticorps monoclonal specifique aux cellules souches embryonnaires humaines
US7923013B2 (en) 2004-12-28 2011-04-12 The Rockefeller University Glycolipids and analogues thereof as antigens for NKT cells
EP1833489A4 (fr) 2004-12-28 2011-08-03 Univ Rockefeller Glycolipides et analogues associes utilises en tant qu'antigenes des lymphocytes nkt
DK1835937T3 (da) 2005-01-06 2012-07-23 Novo Nordisk As Sammensætninger og fremgangsmåder til behandling af virusinfektion
US7837990B2 (en) 2005-03-28 2010-11-23 The Rockefeller University In vivo expanded NKT cells and methods of use thereof
ATE517183T1 (de) 2005-03-31 2011-08-15 Biomedics Inc Monoklonaler antikörper gegen cd20
AU2006250888A1 (en) 2005-05-24 2006-11-30 Avesthagen Limited A method for the production of a monoclonal antibody to CD20 for the treatment of B-cell lymphoma
CN101282993A (zh) 2005-06-02 2008-10-08 阿斯利康公司 针对cd20的抗体和其用途
JP2007036104A (ja) 2005-07-29 2007-02-08 Nec Electronics Corp 半導体装置およびその製造方法
AU2006308847C1 (en) 2005-10-31 2012-05-10 Oncomed Pharmaceuticals, Inc. Compositions and methods for treating and diagnosing cancer
MY149159A (en) 2005-11-15 2013-07-31 Hoffmann La Roche Method for treating joint damage
US7781203B2 (en) 2005-12-29 2010-08-24 Corning Incorporated Supports for assaying analytes and methods of making and using thereof
US20090060921A1 (en) 2006-01-17 2009-03-05 Biolex Therapeutics, Inc. Glycan-optimized anti-cd20 antibodies
WO2007133855A2 (fr) 2006-03-27 2007-11-22 University Of Maryland Biotechnology Institute Synthèse de glycoprotéines et remodelage par transglycosylation enzymatique
PL2018564T3 (pl) 2006-05-18 2010-12-31 Veterinaermedizinische Univ Wien Sposób wykrywania wirusów grypy
WO2008006373A1 (fr) 2006-07-12 2008-01-17 Merck Patent Gmbh Détection en phase solide de monosaccharides terminaux clivés de substrats glycosylés
JP2008025989A (ja) 2006-07-15 2008-02-07 Keio Gijuku 局在表面プラズモン共鳴法と質量分析法によるリガンドの分析方法及びそのためのセンサー素子
EP2059593A2 (fr) 2006-08-18 2009-05-20 Oncotherapy Science, Inc. Traitement ou prévention de cancers surexprimant le reg4 ou le kiaa0101
EP2962697A1 (fr) 2006-11-27 2016-01-06 diaDexus, Inc. Compositions d'anticorps ovr110 et procédés d'utilisation
US8765390B2 (en) 2006-12-08 2014-07-01 The Board Of Trustees Of The Leland Stanford Junior University Identification and isolation of squamous carcinoma stem cells
CA2591496C (fr) 2006-12-18 2014-09-02 Japan Science And Technology Agency Methode de mesure de l'interaction entre un biomateriau et une chaine glucidique, methode d'evaluation de biomateriau dans la selectivite de la chaine glucidique, methode de tri de biomateriau, methode de prise pour modele de biomateriaux, et trousses de mise en oeuvre
WO2008087257A1 (fr) 2007-01-18 2008-07-24 Suomen Punainen Risti, Veripalvelu Nouveaux procédés et réactifs pour la production de cellules
EP2115461A4 (fr) 2007-01-18 2010-01-13 Suomen Punainen Risti Veripalv Nouveaux agents de liaison cellulaire specifiques
US8309354B2 (en) 2007-01-22 2012-11-13 Macrogenics West, Inc. Human cancer stem cells
SI2123271T1 (sl) 2007-03-07 2012-05-31 Daiichi Sankyo Co Ltd Zdravilo za zdravljenje influence
WO2008153615A2 (fr) 2007-03-07 2008-12-18 Ada Technologies, Inc. Microréseau de carbohydrate et nanoparticules conjuguées et procédés de fabrication
US7960139B2 (en) 2007-03-23 2011-06-14 Academia Sinica Alkynyl sugar analogs for the labeling and visualization of glycoconjugates in cells
US7943330B2 (en) 2007-03-23 2011-05-17 Academia Sinica Tailored glycoproteomic methods for the sequencing, mapping and identification of cellular glycoproteins
EP2308514B1 (fr) 2007-03-23 2013-06-05 to-BBB Holding B.V. Conjugés pour le transport de médicaments à travers la barrière hémato-encéphalique
WO2008128207A1 (fr) 2007-04-13 2008-10-23 Academia Sinica Analogues d'alpha-galactosyl céramide et leur utilisation en immunothérapies
JP2010526066A (ja) 2007-04-23 2010-07-29 シェーリング コーポレイション 抗ｍｄｌ−１抗体
EP2527435B2 (fr) 2007-07-12 2021-10-13 Sangamo BioSciences, Inc. Procédés et compositions pour inactiver l'expression génique d'alpha-1,6-fucosyltransférase (FUT 8)
EP2022848A1 (fr) 2007-08-10 2009-02-11 Hubrecht Institut Procédé d'identification, d'expansion, et de suppression de cellules souches adultes et cellules souches de cancer
US20110306049A1 (en) 2007-08-24 2011-12-15 Tokyo Institute Of Technology Method for detecting gynecologic cancer
JP5509080B2 (ja) 2007-08-31 2014-06-04 アカデミア シニカ 抗インフルエンザ活性を有するオセルタミビル含有ホスホネートコンジナーの合成
FR2921387B1 (fr) 2007-09-26 2012-04-20 Sanofi Pasteur Procede de production du virus de la grippe
US20090123439A1 (en) 2007-11-09 2009-05-14 The Jackson Laboratory Diagnostic and prognosis methods for cancer stem cells
BRPI0908968A8 (pt) 2008-03-21 2016-06-28 Danisco Us Inc composição enriquecidas com hemicelulase para potencializar a hidrólise de biomassa
EP2272854B1 (fr) 2008-03-25 2015-08-05 Riken Nouveau glycolipide et son utilisation
MX2010011176A (es) 2008-04-09 2011-02-24 Becton Dickinson Co Inmunoanalisis sensible que utiliza nanoparticulas revestidas.
US8383554B2 (en) 2008-04-14 2013-02-26 Academia Sinica Quantitative microarray of intact glycolipid CD1d interaction and correlation with cell-based cytokine production
US8906832B2 (en) 2008-04-30 2014-12-09 Academia Sinica Quantitative analysis of carbohydrate-protein interactions using glycan microarrays: determination of surface and solution dissociation constants
WO2009140853A1 (fr) 2008-05-23 2009-11-26 The University Of Hong Kong Thérapie de combinaison pour le traitement de la grippe
US20110137570A1 (en) 2008-05-30 2011-06-09 Anthony Lapadula Methods for structural analysis of glycans
NZ590064A (en) 2008-06-16 2012-05-25 Academia Sinica Cancer diagnosis based on levels of antibodies against globo h and its fragments
JP2011524375A (ja) 2008-06-16 2011-09-01 アカデミア シニカ ＧｌｏｂｏＨおよびＳＳＥＡ３に特異的な免疫反応を誘起する組成物および癌治療におけるその使用
JP2010014691A (ja) 2008-06-20 2010-01-21 Igaku Seibutsugaku Kenkyusho:Kk 腹水中のメソテリン及び／又は巨核球増強因子を検出するための方法、キット、試薬及び装置
US20100003674A1 (en) 2008-07-03 2010-01-07 Cope Frederick O Adult stem cells, molecular signatures, and applications in the evaluation, diagnosis, and therapy of mammalian conditions
US7928077B2 (en) 2008-07-11 2011-04-19 Academia Sinica Alpha-galactosyl ceramide analogs and their use as immunotherapies
DK2318832T3 (da) 2008-07-15 2014-01-20 Academia Sinica Glycan-arrays på PTFE-lignende aluminiumcoatede objektglas og relaterede fremgangsmåder
US20100022916A1 (en) 2008-07-24 2010-01-28 Javanbakhsh Esfandiari Method and Apparatus for Collecting and Preparing Biological Samples for Testing
GB0816679D0 (en) 2008-09-11 2008-10-22 Univ Bath Compounds for treating viral infections
JP2012503656A (ja) 2008-09-26 2012-02-09 エウレカ セラピューティクス，インコーポレイテッド 変異体グリコシル化パターンを有する細胞株およびタンパク質
EP3118326B1 (fr) 2008-10-27 2020-04-29 Sapporo Medical University Marqueur moléculaire de cellule souche cancéreuse
WO2010111703A1 (fr) 2009-03-27 2010-09-30 Academia Sinica Donneurs de sialylphosphate alpha-sélectifs pour la préparation de sialosides et puces à sialoside pour la détection de l'influenzavirus
WO2011005756A1 (fr) 2009-07-06 2011-01-13 Puretech Ventures, Llc Administration d'agents ciblés contre des niches de microbiote
WO2011006237A1 (fr) 2009-07-15 2011-01-20 The University Of British Columbia Glycosides 2,3-fluorés en tant qu'inhibiteurs de neuraminidase et leur utilisation en tant qu'agents antiviraux
US20120171201A1 (en) 2009-07-22 2012-07-05 Enzon Pharmaceuticals, Inc. Methods of treating her2 positive cancer with her2 receptor antagonist in combination with multi-arm polymeric conjugates of 7-ethyl-10-hydroxycamptothecin
US20120172329A1 (en) 2009-09-14 2012-07-05 Thailand Excellence Center For Tissue Engineering Phytochemical compositions including xanthones for anti-inflammatory, anti-cytokine storm, and other uses
WO2011074621A1 (fr) 2009-12-18 2011-06-23 株式会社医学生物学研究所 Anticorps contre la mesotheline (msln), et mise en œuvre de celui-ci
EP2347769A1 (fr) 2010-01-20 2011-07-27 Glycotope GmbH Marqueurs de cellules souches de cancer et utilisations associées
JP2013519682A (ja) 2010-02-11 2013-05-30 アレクシオン ファーマシューティカルズ， インコーポレイテッド 抗ｃｄ２００抗体を使用する治療方法
AU2011220878A1 (en) 2010-02-24 2012-08-23 Merck Sharp & Dohme Corp. Method for increasing N-glycosylation site occupancy on therapeutic glycoproteins produced in Pichia pastoris
AR080513A1 (es) 2010-03-12 2012-04-11 Inmunogen Inc Moleculas de union cd37 y sus inmunoconjugados
WO2011130332A1 (fr) 2010-04-12 2011-10-20 Academia Sinica Puces au glycane pour la recherche par criblage haut débit de virus
WO2011130624A2 (fr) 2010-04-16 2011-10-20 Immune Disease Institute, Inc. Expression de polypeptide prolongée à partir d'arn synthétiques modifiés et utilisations de celle-ci
US9403855B2 (en) 2010-05-10 2016-08-02 Academia Sinica Zanamivir phosphonate congeners with anti-influenza activity and determining oseltamivir susceptibility of influenza viruses
WO2011145957A1 (fr) 2010-05-20 2011-11-24 Auckland Uniservices Limited Agents et procédés de détection et/ou d'imagerie d'hypoxie
CN103080130B (zh) 2010-05-27 2016-08-17 默沙东公司 制备具有改进特性的抗体的方法
GB201015569D0 (en) 2010-09-16 2010-10-27 Medical Res Council Blood assay for prions
WO2012082635A1 (fr) 2010-12-13 2012-06-21 Ancora Pharmaceuticals, Inc. Streptocoque du groupe a à base d'oligosaccharides synthétiques
CN103748103A (zh) 2011-01-05 2014-04-23 台湾大学 制备鞘糖脂的方法及其应用
CA2865819A1 (fr) 2011-02-28 2013-09-07 Mcmaster University Traitement du cancer par des antagonistes des recepteurs dopaminergiques
AU2012258907A1 (en) 2011-05-25 2013-11-07 Merck Sharp & Dohme Corp. Method for preparing Fc-containing polypeptides having improved properties
EP3418300B1 (fr) 2011-07-18 2020-10-28 Institute for Research in Biomedicine Anticorps neutralisant le virus de la grippe a et leurs utilisations
KR102021159B1 (ko) 2011-08-12 2019-09-11 닛산 가가쿠 가부시키가이샤 트리시클릭 헤테로시클릭 화합물 및 jak 저해제
CA2853809A1 (fr) 2011-10-31 2013-05-10 Merck Sharp & Dohme Corp. Procede de preparation d'anticorps possedant des proprietes ameliorees
WO2013074598A1 (fr) 2011-11-18 2013-05-23 Merck Sharp & Dohme Corp. Polypeptides contenant fc ayant des propriétés anti-inflammatoires améliorées et une liaison améliorée à fcrn
EP2604281B1 (fr) 2011-12-14 2014-07-30 Centre National de la Recherche Scientifique (CNRS) Analogues conjugués de somatostatine coupés pour des applications biologiques
US9637465B2 (en) 2012-01-19 2017-05-02 The University Of British Columbia 3′ Equatorial-fluorine-substituted neuraminidase inhibitor compounds, compositions and methods for the use thereof as anti-virals
GB201201314D0 (en) 2012-01-26 2012-03-07 Isis Innovation Composition
EP2812442B9 (fr) * 2012-02-10 2023-02-15 University of Maryland, Baltimore Glyco-ingénierie chimio-enzymatique d'anticorps et de leurs fragments fc
WO2013130603A1 (fr) 2012-02-27 2013-09-06 Board Of Regents, The University Of Texas System Ganglioside gd2 en tant que marqueur et cible sur des cellules souches cancéreuses
JP2015514113A (ja) 2012-04-02 2015-05-18 メリマック ファーマシューティカルズ インコーポレーティッド 単一特異性および二重特異性抗ｉｇｆ−１ｒおよび抗ｅｒｂｂ３抗体の用法および用量
WO2013151649A1 (fr) 2012-04-04 2013-10-10 Sialix Inc Composés d'interaction avec des glycanes
US10130714B2 (en) 2012-04-14 2018-11-20 Academia Sinica Enhanced anti-influenza agents conjugated with anti-inflammatory activity
EP2885311B1 (fr) 2012-08-18 2020-01-01 Academia Sinica Sondes perméables aux cellules pour l'identification et l'imagerie de sialidases
TWI563092B (en) 2012-08-20 2016-12-21 Academia Sinica Large scale enzymatic synthesis of oligosaccharides
US9547009B2 (en) 2012-08-21 2017-01-17 Academia Sinica Benzocyclooctyne compounds and uses thereof
KR102460297B1 (ko) 2012-10-30 2022-10-28 에스퍼란스 파마슈티컬스, 인코포레이티드 항체/약물 컨쥬게이트 및 이의 사용 방법
US20150284452A1 (en) 2012-11-13 2015-10-08 Iogenetics, Llc Antimicrobial compositions
GB201305986D0 (en) 2013-04-03 2013-05-15 Asociaci N Ct De Investigaci N Cooperativa En Biomateriales Synthesis and use of isotopically-labelled glycans
US20160058886A1 (en) 2013-04-13 2016-03-03 Universidade De Coimbra Platform for targeted delivery to stem cells and tumor cells and uses thereof
CN104225616A (zh) 2013-06-08 2014-12-24 中南大学 一种靶向卵巢癌干细胞的抗肿瘤生物制剂
US10086054B2 (en) 2013-06-26 2018-10-02 Academia Sinica RM2 antigens and use thereof
EP3013347B1 (fr) 2013-06-27 2019-12-11 Academia Sinica Conjugués de glycane et leur utilisation
TWI599370B (zh) 2013-07-26 2017-09-21 中央研究院 靈芝多醣誘發之抗體介導抗腫瘤活性
CN105682666B (zh) 2013-09-06 2021-06-01 中央研究院 使用醣脂激活人类iNKT细胞
EP3043647A4 (fr) 2013-09-12 2017-05-10 Teva Pharmaceutical Industries Ltd. Expression génétique de biomarqueurs de la réceptivité au laquinimod
CN103436627B (zh) 2013-09-13 2016-02-03 四川大学华西医院 一种恶性乳腺癌干细胞的筛查试剂盒
WO2015054039A1 (fr) 2013-10-08 2015-04-16 Merck Sharp & Dohme Corp. Polypeptides contenant fc présentant une liaison accrue à fcgammariib
EP3094352B1 (fr) 2014-01-16 2020-09-23 Academia Sinica Compositions et méthodes pour traiter et détecter des cancers
US10150818B2 (en) 2014-01-16 2018-12-11 Academia Sinica Compositions and methods for treatment and detection of cancers
TWI687428B (zh) 2014-03-27 2020-03-11 中央研究院 反應性標記化合物及其用途
US10005847B2 (en) 2014-05-27 2018-06-26 Academia Sinica Anti-HER2 glycoantibodies and uses thereof
EP3904388A1 (fr) 2014-05-27 2021-11-03 Academia Sinica Fucosidase issue de bacteroïdes et ses procédés d'utilisation
AU2015267047A1 (en) 2014-05-27 2017-01-05 Academia Sinica Anti-CD20 glycoantibodies and uses thereof
US10118969B2 (en) 2014-05-27 2018-11-06 Academia Sinica Compositions and methods relating to universal glycoforms for enhanced antibody efficacy
WO2015184001A1 (fr) 2014-05-28 2015-12-03 Academia Sinica Glycoanticorps anti-thf-alpha et leurs utilisations
KR102003138B1 (ko) 2014-08-22 2019-07-23 아카데미아 시니카 신규한 글리칸 접합체 및 그의 용도
KR102422375B1 (ko) 2014-09-08 2022-07-18 아카데미아 시니카 당지질을 사용한 인간 iNKT 세포 활성화
CN114106179A (zh) 2014-09-12 2022-03-01 加利福尼亚大学董事会 巨胞饮人类抗cd46抗体和靶向癌症疗法
US9975965B2 (en) 2015-01-16 2018-05-22 Academia Sinica Compositions and methods for treatment and detection of cancers
US10495645B2 (en) 2015-01-16 2019-12-03 Academia Sinica Cancer markers and methods of use thereof
WO2016118090A1 (fr) 2015-01-23 2016-07-28 Agency For Science, Technology And Research Protéines de liaison à l'antigène spécifiques du cancer
AU2015378564A1 (en) 2015-01-24 2017-07-13 Academia Sinica Novel glycan conjugates and methods of use thereof
US20170283878A1 (en) 2015-12-11 2017-10-05 Academia Sinica Modulation of globoseries glycosphingolipid synthesis and cancer biomarkers
WO2017156192A1 (fr) 2016-03-08 2017-09-14 Academia Sinica Procédés de synthèse modulaire de n-glycanes et de puces à n-glycanes
BR112018070097A2 (pt) 2016-03-29 2019-02-12 Obi Pharma, Inc. anticorpo, hibridoma, composição farmacêutica, método para tratar câncer em um indivíduo, método para inibir a proliferação de células cancerígenas, método para diagnosticar o câncer em um indivíduo, método para tratar um paciente humano, método para fazer imagens de um indivíduo, conjugado de fármaco-anticorpo (adc), método para tratar câncer, anticorpo biespecífico e método para preparar uma população de anticorpos homogêneos
JP7213549B2 (ja) 2016-08-22 2023-01-27 シーエイチオー ファーマ インコーポレイテッド 抗体、結合性断片、および使用の方法
WO2018094414A1 (fr) 2016-11-21 2018-05-24 Obi Pharma, Inc. Molécules biologiques conjuguées, compositions pharmaceutiques et procédés

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6528286B1 (en) * 1998-05-29 2003-03-04 Genentech, Inc. Mammalian cell culture process for producing glycoproteins
US20060073161A1 (en) * 1999-04-09 2006-04-06 Breton Gary L Nucleic acid and amino acid sequences relating to Bacteroides fragilis for diagnostics and therapeutics
US20100173323A1 (en) * 2006-06-09 2010-07-08 University Of Maryland, Baltimore Glycosylation engineered antibody therapy
US20130337018A1 (en) * 2007-10-02 2013-12-19 Avaxia Biologics, Incorporated Antibody therapy for use in the digestive tract
US20130230886A1 (en) * 2007-12-31 2013-09-05 Christian Votsmeier Antibodies to TNF Alpha
US20110263828A1 (en) * 2009-12-02 2011-10-27 Academia Sinica Methods for modifying human antibodies by glycan engineering
US20120226024A1 (en) * 2011-03-03 2012-09-06 Lai-Xi Wang Core fucosylated glycopeptides and glycoproteins: chemoenzymatic synthesis and uses thereof
WO2013181585A2 (fr) * 2012-06-01 2013-12-05 Momenta Pharmaceuticals, Inc. Méthodes associées à l'adalimumab

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
See also references of EP3154582A4 *
TSAI ET AL.: "Effective Sugar Nucleotide Regeneration for the Large-Scale Enzymatic Synthesis of Globo H and SSEA4", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 135, no. 39, 17 September 2013 (2013-09-17), pages 14831 - 14839, XP055218268, ISSN: 0002-7863 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3257866A1 (fr) * 2016-06-17 2017-12-20 Academisch Medisch Centrum Anticorps anti-tnf modifiés et leur utilisation dans le traitement de la maladie intestinale inflammatoire
WO2019234021A1 (fr) * 2018-06-04 2019-12-12 Limmatech Biologics Ag Anticorps monoclonal glycomodifié

Also Published As

Publication number Publication date
TW201609135A (zh) 2016-03-16
JP2020176144A (ja) 2020-10-29
IL249184B (en) 2022-04-01
US20150344559A1 (en) 2015-12-03
EP3154582A1 (fr) 2017-04-19
TWI732738B (zh) 2021-07-11
US11332523B2 (en) 2022-05-17
KR20170004023A (ko) 2017-01-10
JP7063538B2 (ja) 2022-05-09
CA2950433A1 (fr) 2015-12-03
CN106714829A (zh) 2017-05-24
WO2015184001A8 (fr) 2021-10-21
IL249184A0 (en) 2017-01-31
AU2015267044A1 (en) 2016-12-15
EP3154582A4 (fr) 2018-01-10
JP2017518990A (ja) 2017-07-13
KR102494193B1 (ko) 2023-01-31

Similar Documents

Publication Publication Date Title
US11332523B2 (en) Anti-TNF-alpha glycoantibodies and uses thereof
US10618973B2 (en) Anti-HER2 glycoantibodies and uses thereof
US11884739B2 (en) Anti-CD20 glycoantibodies and uses thereof
EP3149045B1 (fr) Compositions et procédés concernant des glycoformes universelles pour une efficacité d'anticorps améliorée
US11377485B2 (en) Methods for modifying human antibodies by glycan engineering

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15800059

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 249184

Country of ref document: IL

ENP Entry into the national phase

Ref document number: 2950433

Country of ref document: CA

Ref document number: 2016569732

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

REEP Request for entry into the european phase

Ref document number: 2015800059

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2015800059

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2015267044

Country of ref document: AU

Date of ref document: 20150527

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20167036499

Country of ref document: KR

Kind code of ref document: A